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Topic:Fertility

Fertility in horses encompasses the physiological processes and factors influencing reproductive success in equine species. It involves the study of reproductive anatomy, endocrinology, and behavior, as well as the management practices that affect breeding outcomes. Key aspects include the estrous cycle, conception rates, and factors impacting stallion and mare fertility. Reproductive technologies such as artificial insemination, embryo transfer, and hormonal therapies are also explored to enhance breeding efficiency. This page compiles peer-reviewed research studies and scholarly articles that examine the biological mechanisms, management strategies, and technological advancements related to equine fertility.
Seminal plasma exposure modulates peripheral T cell-associated transcription during early pregnancy in the mare.
Veterinary immunology and immunopathology    July 21, 2026   Volume 300 111179 doi: 10.1016/j.vetimm.2026.111179
Crook RA, Branch AE, Kapadia CL, Klinglesmith BA, Hatzel JN, Sones JL, Fedorka CE.Successful establishment of pregnancy requires maternal immune tolerance to paternal antigens introduced by insemination. Regulatory T cell (Treg)-associated responses are thought to play a central role in this process, whereas pro-inflammatory T helper 1 (Th1) responses may disrupt pregnancy success. In other species, seminal plasma (SP) has been shown to modulate maternal immune adaptation; however, this relationship remains poorly defined in mares, where SP is frequently reduced during routine breeding practices. The objectives of this study were to (1) characterize temporal changes in syst...
Density gradient centrifugation preserves sperm kinetics and morphology in stallion ejaculates with severe urospermia.
Journal of equine veterinary science    July 18, 2026   Volume 166 106080 doi: 10.1016/j.jevs.2026.106080
Miranda TGR, Baldini PHO, Silva AC, Rebelo LS, Oliveira AJS, Ladislau PT, Ignácio FS, Papa FO, Monteiro GA.Urospermia is a frequent cause of subfertility in stallions due to the rapid deterioration of sperm quality. Objective: This study aimed to evaluate the effectiveness of different processing methods in minimizing the deleterious effects of urine on cooled equine semen. Methods: Ejaculates from 12 stallions were subjected to simple dilution, conventional centrifugation, and colloidal density gradient centrifugation, under challenges of 0%, 20%, and 50% urine, and stored at 5 °C for 24 h. Results: A severe dose-dependent effect was observed: increasing urinary concentration progressively reduce...
Sequencing and Differential Analysis of miRNA in the Ovaries of Kazakh Horses During Estrus and Anestrus States.
Animals : an open access journal from MDPI    July 15, 2026   Volume 16, Issue 14 2197 doi: 10.3390/ani16142197
Liu J, Yao X, Meng J, Wang J, Zeng Y, Ren W.To reveal the regulatory role of miRNA in the estrous cycle transition of Kazakh horse ovaries and improve their reproductive efficiency, this study took the ovaries of Kazakh horses in the estrus period (DY group) and the quiescent period (DB group) as research objects. Small RNA sequencing and bioinformatics analysis were used to systematically analyze the expression characteristics and functional pathways of miRNA under the two physiological states. A total of 161 million high-quality miRNA tags were obtained through sequencing. Principal component analysis showed that the intra-group repea...
Isolation of high-quality stallion spermatozoa: a comparison of methods and their effects on sperm quality and yield, DNA damage, and functional competence by ICSI.
Reproduction, fertility, and development    July 15, 2026   Volume 38, Issue 11 RD25242 doi: 10.1071/RD25242
Bahrami M, Medica AJ, Clutton-Brock A, Herzog F, Gibb Z, Bakos HW, Aitken RJ, Swegen A.Intracytoplasmic sperm injection (ICSI) is the primary method for equine in vitro embryo production, yet outcomes may be influenced by sperm selection techniques. Objective: This study compared four isolation methods: EquiPure™, Vetmotl™, Felix™, and Samson™, to assess effects on sperm quality and embryo development. Methods: Semen was collected from four fertile stallions, processed and evaluated for sperm motility, viability, DNA integrity, and yield of high-quality cells relative to input. Isolated samples were compared to non-isolated (centrifuged) samples. Spermatozoa were used fo...
Pre-ovulatory follicles and ipsilateral ovarian stroma are cooler than non-dominant antral follicles and stroma on the contralateral ovary of the mare.
Theriogenology    July 14, 2026   Volume 265 118081 doi: 10.1016/j.theriogenology.2026.118081
Hoogewijs M, Bussade P, Ismer A, Grippo A, Santana B, Wilsher S.Reports in several species have demonstrated that pre-ovulatory follicles (POFs) are cooler than neighbouring ovarian tissue, with cooling important for fertility. This study measured the temperature of POFs and compared it with other regions of the ovary, cervix, and rectum to determine if POF cooling occurs in mares. Fourteen oestrous cycles in 11 mares were monitored, and when an ovarian follicle ≥38 mm was present, deslorelin was given. Thirty hours later, rectal and cervical temperature measurements were taken. Mares were sedated and ovarian temperature measurements taken using a ther...
Sperm Energy-restriction and Recovery (SER) enhances equine sperm function and embryo development after ICSI.
Journal of animal science    July 10, 2026   skag213 doi: 10.1093/jas/skag213
Arroyo-Salvo C, Rio S, Bogetti ME, Clerico G, Morado S, Turner R, Visconti PE, Gambini A, Perez-Martinez S, Gervasi MG.Assisted reproductive technologies (ART) are extensively used in domestic animal reproduction; however, improving their efficiency and consistency remains a major challenge for achieving reliable reproductive outcomes. The equine industry faces unique challenges related to sperm capacitation, which limited the success of conventional in vitro fertilization (IVF) for many years. Consequently, intracytoplasmic sperm injection (ICSI) became the most widely used technique for commercial in vitro embryo production. Even though a successful IVF protocol was recently developed, substantial opportunit...
Factors Influencing Equine Embryo Outcomes in OPU‑ICSI Systems: Insights from a Large‑Scale Program in Brazil.
Journal of equine veterinary science    July 6, 2026   106073 doi: 10.1016/j.jevs.2026.106073
Bordignon V, Buratini J, Cue RI, Barrilari V, Gonçalves MA, Merlini G, Fernandes CB, Squires E, Pontes JHF, Fleury PDC.Equine OPU‑ICSI supports commercial IVEP, yet substantial variability persists. We retrospectively analyzed a large dataset generated in Brazil spanning Quarter Horses, Warmbloods, and Mangalarga to evaluate whether predictors previously identified in the literature show comparable patterns under distinct ecological and breeding conditions. Overall oocyte and embryo outcomes were aligned with broadly reported trends from previous studies. Oocytes retrieved from estrous mares bearing a dominant follicle exhibited higher maturation and blastocyst rates. Stallion effects segregated into distinc...
Blood ammonia levels: A potential indicator of high-risk pregnancy in Thoroughbred broodmares.
Veterinary and animal science    June 25, 2026   Volume 34 100750 doi: 10.1016/j.vas.2026.100750
Taylor JR, Dorton AR, Robertson SL.Exposure to high blood ammonia (hyperammonemia) interferes with gestation due to its detrimental effect on hormone levels, fetal development, the immune system, placental integrity, and fertility of broodmares. Increases in ammonia levels are dangerous and elusive, as damage may occur from earlier peaks, resulting in the trapping of ammonia in the amniotic fluid and then disappearing from the blood circulation before clinical symptoms are observed. Thus, it should be recognized early and treated immediately before complications arise. This study aimed to investigate if blood ammonia levels in ...
Advanced paternal age is associated with reduced reproductive success and offspring racing performance in Australian Thoroughbred racehorses.
Scientific reports    June 23, 2026   doi: 10.1038/s41598-026-59342-9
Jenkins C, Upton R, Griffin RA, Swegen A, Hamilton NA, Gibb Z.Reproductive ageing influences fertility and offspring fitness across mammals. In humans and model organisms, paternal age has been associated with changes in sperm quality and genomic integrity, including oxidative stress, DNA damage, and epigenetic alterations; however, these mechanisms have not been directly examined in large equine populations. Thoroughbred racehorses provide a system to investigate paternal age effects at scale using detailed breeding records. In this retrospective observational study, with data from 35 retired stallions and 33,546 progeny, we assessed associations betwee...
Matched embryo-endometrium RNA-seq reveals coordinated but asymmetric transcriptomic reprogramming at the onset of early equine pregnancy.
Frontiers in cell and developmental biology    June 18, 2026   Volume 14 1840498 doi: 10.3389/fcell.2026.1840498
Da Silva Álvarez E, Ortiz-Rodríguez JM, Martín-Cano FE, Álvarez-Barrientos A, Becerro-Rey L, Gil MC, Redondo E, Masot J, Aparicio IM, Peña FJ....Maternal recognition of pregnancy (MRP) in the mare requires coordinated adaptations of both the conceptus and the endometrium during the peri-recognition interval; however, integrated transcriptomic analyses of both compartments within the same biological window remain limited. Here, we analyzed conceptus RNA-seq data across embryonic age (Days 8 and 12 post ovulation) and endometrial RNA-seq data across pregnancy status (pregnant versus non-pregnant mares sampled between Days 8 and 12 post ovulation). In pregnant mares, conceptus recovery and endometrial biopsy collection were performed with...
Redefining stallion sperm bioenergetics: lessons from a decade of change in mammalian sperm metabolism.
Reproduction (Cambridge, England)    June 17, 2026   Volume 172, Issue 1 xaag077 doi: 10.1093/reprod/xaag077
Peña FJ, Becerro-Rey L, Ortega-Ferrusola C, da Silva-Álvarez E, Zabalo-Palomo C, Gil MC, Martín-Cano FE.In brief: Sperm bioenergetics is now understood as an integrated system in which ATP production, redox homeostasis, substrate availability, and cellular heterogeneity jointly determine sperm function. This review uses the stallion as an oxidative-biased model to synthesize advances from 2016 to 2026 and proposes a metabolically plastic framework with direct implications for semen preservation and fertility assessment. Abstract: Over the last decade, mammalian sperm metabolism has moved from a binary view centered on glycolysis versus oxidative phosphorylation to an integrated model in which AT...
Achieving Success in Conventional Equine Embryo Transfer: A Practical Review.
Journal of equine veterinary science    June 17, 2026   106065 doi: 10.1016/j.jevs.2026.106065
Necchi D, Donato GG, Vandaele H.Equine embryo transfer is a well-established procedure in horse breeding programs and it's used to increase the number of offspring from valuable mares, obtain embryos from mares in competition and produce foals from mares unable to carry a pregnancy to term. The success of embryo recovery depends mainly on the day of flushing, management of insemination, and the fertility of donor mare and stallion. Other relevant factors include donor age and reproductive status, number of ovulations, operator experience, and flushing technique. Pregnancy rates after transfer are influenced by donor-recipien...
Chromosomal chaos: understanding aneuploidy associated with assisted reproductive technologies.
Journal of equine veterinary science    June 17, 2026   Volume 164 106053 doi: 10.1016/j.jevs.2026.106053
McKane AM, Chauhan IV, Miller D, Ghosh S, Das A, de Mestre AM.Pregnancy loss remains a significant challenge for the equine breeding industry, with substantial economic and management consequences. Chromosomal abnormalities have emerged as a central driver of pregnancy loss, particularly during early gestation, highlighting the need to better understand the etiology of this inherent genetic instability. Concurrently, the rapid expansion of assisted reproductive technologies (ART) offers promise for improving equine reproductive efficiency alongside the traditional practice of natural cover. However, the increasing use of in vitro embryo production (IVP) ...
Effect of Spirulina platensis extract (C-phycocyanin) supplementation to the semen extender on kinematic and oxidative parameters of Marwari stallions during freezing.
Tropical animal health and production    June 12, 2026   Volume 58, Issue 5 326 doi: 10.1007/s11250-026-05136-0
Yadav V, Talluri TR, Dholpuria S, Khandelwal S, Joshi A.The current research aimed at examining the impact of different quantities of Spirulina platensis extract (C-phycocyanin) supplemented to the semen extender on the post-thaw quality of Marwari stallion spermatozoa. Semen samples were obtained via an artificial vagina from six fertile stallions and diluted with an extender containing Spirulina platensis extract (C-phycocyanin) at concentrations of 10, 20, and 50 µg/mL, or without Spirulina as a control. Aliquots of diluted semen were subjected to cooling (4 °C, 120 min) and then frozen (-196 °C, 1 day), after which the kinematic paramet...
Cracks in the ice: biological challenges and incremental progress in equine oocyte vitrification.
Journal of equine veterinary science    June 11, 2026   106059 doi: 10.1016/j.jevs.2026.106059
Serrano-Revuelta E, Ángel-Vélez D, Van Soom A, Smits K.Equine oocyte vitrification could enhance the flexibility of in vitro embryo production, yet developmental competence remains reduced. Only vitrification of oocytes matured in vivo yields foaling rates comparable to fresh oocytes. Recent advances in vitrification at the germinal vesicle (GV) stage, supported by systematic comparisons of cryoprotective agents and exposure times, has enabled consistent blastocyst production and birth of several foals. Comparative studies of GV and metaphase II (MII) stages suggest biological advantages of vitrification after in vitro maturation (IVM), warranting...
Cytogenetic, genomic, and reproductive characterization of four pura Raza Española mares carrying a 64, X, del(X),t(X; Y) complex karyotype.
Journal of animal science    June 9, 2026   skag185 doi: 10.1093/jas/skag185
Arjona-Delgado MY, Pirosanto Y, Raudsepp T, Vargas-Pérez MÁ, Valera M, Encina A, Azor P, Bugno-Poniewierska M, Demyda-Peyrás S.Chromosomal abnormalities involving the sex chromosomes constitute critical genetic factors influencing fertility in horses. Nevertheless, a substantial proportion of individuals carrying sex chromosomal abnormalities exhibit normal reproductive phenotypes, which complicates their identification until adulthood. Here, we present four mares carrying a complex deletion/translocation rearrangement with the karyotype formula 64, X, del(X)(p∼25),t(X; Y)(p∼25; q∼15). The cases analyzed belong to three unrelated families of Pura Raza Española (PRE), all presenting the same chromosomal abnormal...
Novel technique for correcting urovagina via urethral transposition in the horse.
Equine veterinary journal    June 7, 2026   doi: 10.1002/evj.70210
McCarthy R, Kamm JL, Pierce M, Huber M.Vesicovaginal reflux is a common cause of infertility in mares. Surgical correction is commonly recommended and involves caudal relocation of the urethral orifice which forces the urine to be expelled caudally. There have been several previously described techniques. Complications associated with caudal urethral extension occur in 11-33.3% of cases. Objective: To identify a novel technique for performing caudal urethral translocation that effectively corrects vesicovaginal reflux with minimal associated complications. Methods: In vivo experiment and clinical case series. Methods: The experimen...
Annual Variation in Stallion Testicular Measurements and Its Association With Hormonal Profiles and Semen Quality.
Reproduction in domestic animals = Zuchthygiene    June 3, 2026   Volume 61, Issue 6 e70244 doi: 10.1111/rda.70244
Braga TRC, Freitas ML, Martins DMV, Viu MAO, Oliveira RA.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) ...
Application of Anti-Müllerian Hormone in Regulation of Ovarian Function and Its Practical Relevance for Fertility and Embryo Production in Cows and Mares.
Veterinary sciences    June 3, 2026   Volume 13, Issue 6 552 doi: 10.3390/vetsci13060552
Vernunft A, Scarlet D.In the female organism, Anti-Müllerian hormone (AMH) is produced exclusively by granulosa cells, particularly in secondary and early tertiary follicles. The amount of AMH released into the blood is proportional to the total number of follicles present in the ovaries and is therefore used to estimate the ovarian functional reserve. In both cattle and mares, a strong and highly repeatable association has also been demonstrated between AMH concentrations and antral follicle count. Therefore, AMH may serve as a biomarker with broad diagnostic utility in reproductive medicine in these species, as ...
Ovum pick-up in mares: Evidence review and best practice recommendations using a modified GRADE approach.
Theriogenology    June 2, 2026   Volume 264 118021 doi: 10.1016/j.theriogenology.2026.118021
Lewis N, Dini P, Spanner J, Claes A, De Grauw J, Morganti M, Daels P, Cuervo-Arango J.Ovum pick-up (OPU) is a procedure that is now performed in mares worldwide to harvest oocytes for in vitro embryo production. There is a lack of consensus surrounding best practices for various procedural elements of the technique and recently there has been an increasing focus on the welfare aspects of the procedure. Our objective was to summarize current practice and to develop guidelines for OPU in the mare, using a modified GRADE approach to optimize efficiency and mare welfare. Published evidence on various aspects of the procedure was compiled, and to supplement the relative lack of publ...
Vitrification of in vivo produced equine embryos: Size really does matter!
Journal of equine veterinary science    June 1, 2026   Volume 164 106032 doi: 10.1016/j.jevs.2026.106032
Wilsher S.Vitrification rather than slow-freezing has become the method of choice for the cryopreservation of in-vivo produced embryos. It is a simple technique that does not require a programmable freezing machine and relies chiefly on placing the embryo on a surface, rather than within a straw, in a small volume of vitrification solution to achieve extremely rapid cooling rates. However, in the field of equine embryo vitrification size truly does matter. Methodologies for both vitrification and warming prior to transfer are largely dictated by the size of the embryo. Since embryo size is directly rela...
Embryo developmental kinetics and vitrification in relation to pregnancy outcome, foaling percentage and foal sex.
Equine veterinary journal    May 31, 2026   doi: 10.1002/evj.70197
Peere S, Dewulf M, Van den Branden E, Papas M, Broothaers K, De Coster T, Briote I, Hedia M, Angel-Velez D, Hoogewijs M, Govaere J, Smits K.While fast in vitro embryo development has been associated with improved outcomes, the relationships of slow in vitro embryo development and vitrified versus fresh in vitro produced (IVP) embryo transfer to foaling percentage and foal sex are less explored. Objective: To determine the relationships of (1) day of blastocyst formation (D7-11 after intracytoplasmic sperm injection (ICSI)) and (2) vitrification and warming method (one- and three-step) versus fresh transfer with pregnancy, early pregnancy loss (EPL), foaling percentage and foal sex. Methods: Retrospective clinical study. Methods: B...
Effect of N-acetyl cysteine treatment on uterine cytokine and chemokine profiles in mares with persistent breeding-induced endometritis.
Journal of equine veterinary science    May 27, 2026   Volume 165 106033 doi: 10.1016/j.jevs.2026.106033
Khan FA, Caissie MD, Wagner B, Scholtz EL, Chenier TS.Persistent breeding-induced endometritis (PBIE) is a major cause of infertility in mares. Recent research has shown that PBIE-affected mares treated with intrauterine N-acetyl cysteine (NAC) 12 hours before breeding showed no improvement in clinical signs at 12- and 60-hours post-breeding. Objective: The present study was designed to examine the effects of NAC treatment on the uterine inflammatory cytokine and chemokine profiles in PBIE-affected mares. Methods: Using a randomized, blinded, cross-over design, mares susceptible to PBIE (n=9) were allocated to Control and Treatment cycles with at...
Comparison of cost for conventional embryo transfer versus intracytoplasmic sperm injection.
Journal of equine veterinary science    May 20, 2026   Volume 164 105937 doi: 10.1016/j.jevs.2026.105937
Squires EL, Ferris R.There are obvious practical advantages of both conventional ET and ICSI. One question that has not been addressed adequately, however, is the relative cost of each procedure and if there is a cost advantage of one over the other. Objective: The objective of the present study was to obtain additional cost data from clinics and breeding farms that perform both ET and ICSI. Methods: A request was sent to 10 clinics that performed either just OPU or OPU/ICSI. These facilities were representative of reproductive specialists located all across the United States. Six clinics provided information on a...
Effects of supplementing concentrates with different energy levels on growth performance, blood indicators, and fecal microbiota of Mongolian horses in late gestation.
BMC veterinary research    May 19, 2026   doi: 10.1186/s12917-026-05568-8
Liu Y, Wang X, Wang G, Wu Z, He Q, Zhao B, Du M, Zhao Y, Bou G, Bai D, Dugarjaviin M, Zhang X.Horse breeding and management have a globally extensive history, with horses serving vital roles in transportation, warfare, agriculture, and cultural activities. In China, particularly in Inner Mongolia, the horse industry faces challenges such as harsh winters and limited nutritional research for local breeds, especially during late gestation. This critical period is essential for fetal development and maternal health, and inadequate nutrition can lead to complications like nutritional abortions and poor foal growth. Similar issues exist worldwide in cold climates or regions with seasonal pa...
Conventional IVF in the horse past, present, and future: physiological and technical insights.
Journal of equine veterinary science    May 18, 2026   105938 doi: 10.1016/j.jevs.2026.105938
Hinrichs K.For decades, conventional in vitro fertilization (IVF) in the horse was ineffective, leading to reliance on intracytoplasmic sperm injection (ICSI) for both research and clinical in vitro embryo production. However, the ability to achieve IVF is essential to the study of key reproductive processes, including sperm capacitation, fertilization, and early embryonic development. Recently we have established a repeatable and efficient IVF system for horses, using long-term (9-22 h) sperm pre-incubation. For clinical use, IVF has many features that may support higher embryo quality than does ICSI, a...
Survival and fertility of Thoroughbred broodmares with uterine tears treated surgically: 76 cases (2000-2023).
Journal of the American Veterinary Medical Association    May 8, 2026   Volume 264, Issue 10 1341-1348 doi: 10.2460/javma.26.01.0050
Nugent C, Embertson R, McQuerry K, Woodie JB, Hopper S.To describe the clinical presentation, treatment, short- and long-term outcomes, and subsequent fertility of Thoroughbred broodmares undergoing surgical repair of periparturient uterine tears. Unassigned: A retrospective observational study was conducted with medical records from a single tertiary referral equine hospital (2000 to 2023). Thoroughbred broodmares with confirmed periparturient uterine tears that underwent surgical repair were included; mares euthanized prior to surgery or intraoperatively or that had incomplete medical records were excluded. Data collected included signalment, cl...
FGF2, LIF, and IGF1 supplementation during in vitro maturation or culture improves equine ICSI embryo development in an age-dependent manner.
Reproduction, fertility, and development    May 7, 2026   Volume 38, Issue 7 RD26023 doi: 10.1071/RD26023
Smith JM, Palacios PD, Segura Forero PA, Gurkin RJ, Doyle T, Boe-Hansen G, Gambini A.Ovum pickup (OPU) combined with intracytoplasmic sperm injection (ICSI) has reshaped equine assisted reproduction, enabling embryo production from mares and stallions with constrained breeding opportunities; however, limited efficiency of in vitro maturation (IVM) and in vitro culture (IVC) continues to hinder embryo production. Growth factor supplementation with fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF1) in combination (FLI) has been shown to enhance oocyte and embryo developmental competence in several species. Objective: This...
The Equine Reproductive Microbiota: Composition, Dynamics, Dysbiosis, and Implications for Fertility in Mares and Stallions.
Animals : an open access journal from MDPI    May 5, 2026   Volume 16, Issue 9 1414 doi: 10.3390/ani16091414
Pelenė U, Šiukščius A, Nainienė R, Merkelytė I, Šveistienė R.The equine reproductive microbiota has emerged as an important factor influencing reproductive health and fertility in both mares and stallions. Traditionally, the equine uterus was considered sterile, and microbial presence was interpreted primarily in the context of infection. However, sequencing-based studies have demonstrated that the reproductive tract, including the uterus and semen, contains detectable microbial communities or microbial DNA signatures, challenging this traditional paradigm. In mares, the vaginal microbiota is consistently dominated by , , , and , whereas the uterine env...
The Effects of L-Citrulline on the Apparent Digestion and Metabolism of Nutrients, Blood Hormone Levels, Amino Acid Metabolism and the Diversity of Faecal Microbiota in Mares in the Later Stage of Pregnancy.
Life (Basel, Switzerland)    April 30, 2026   Volume 16, Issue 5 744 doi: 10.3390/life16050744
Liu P, Yang F, Li J, Li C, Guo X, Li X.This study aimed to investigate the impact of dietary L-citrulline supplementation on the health of mares during late gestation. Thirty-two healthy mares in late pregnancy were randomly assigned to four groups: a control group (CON, 0 g/d) and three treatment groups receiving 15, 30, and 45 g/d/head of L-citrulline, respectively. The trial spanned 72 days, including a 12-day adaptation phase and a 60-day formal feeding period. A fixed daily feeding amount of 11.2 kg/head was provided, ensuring complete consumption and consistent dry matter intake across all groups. Results demonstrated that su...