The objective of the present study was to evaluate the effects of a novel Inhibin (INH) DNA vaccine (C500/pVAX-asd-IS) on the immune response, reproductive hormone levels, and spermatogenesis of rats. Forty healthy male rats were divided into four groups, and respectively immunized (thrice, 14 d apart) with 1×10, 1×10, and 1×10 CFU of the recombinant inhibin vaccine (group C500/pVAX-asd-IS-L, C500/pVAX-asd-IS-M, and C500/pVAX-asd-IS-H) or 1×10 CFU C500. P/N values increased after vaccination and differed ( <0.05) at 7 d, and sharply increased at 14 d following the booster vaccination ( <0.01); The weight and volume of testes in C500/pVAX-asd-IS groups were increased ( < 0.05) at decapitation, respectively; Histological evaluation showed that the number of spermatogenic cells in the lumen was increased, and the cytoplasmic remnants of sperms were allergy increased significantly compared with the control group. Oral vaccination with INH DNA reduced ( < 0.05) serum concentrations of INH B, enhanced serum concentrations of testosterone (T) and FSH. Furthermore, mRNA expressions of and in the testes were increased in C500/pVAX-asd-IS-M and C500/pVAX-asd-IS-H groups ( < 0.05 or < 0.01). The mRNA amount of in C500/pVAX-asd-IS-M group was greater than control group ( < 0.05).These results suggested that neutralization of endogenous INH through oral vaccination with INH DNA delivered by C500 strain successfully elicited a humoral immune response. INH gene immunization may have a positive effect on spermatogenesis and reproductive efficiency in male rats.

The path to fertility in the mare requires an understanding of the hormonal influences, the immune response, genetics, and epigenetic mechanisms involved not only in physiological reproductive processes, but also such pathologies as endometritis and endometrosis. Endometritis may lead to endometrosis establishment. In the presence of endometritis, neutrophils arrive at the mare endometrium, and form neutrophil extracellular traps. While NETosis plays pivotal roles, prolonged inflammation can lead to chronic endometritis, endometrosis, and fertility issues. Matrix metalloproteinases and epigenetic changes influence the course of endometrosis. Inhibitors of specific enzymes involved in NETosis and epigenetic inhibitors have shown potential in reducing pro-fibrotic effects. Collagen type III (COL3) has emerged as a putative biomarker, correlating with endometrosis and useful in fertility assessment. Thus, COL3 may offer a non-invasive diagnostic tool, as a complement to histopathological methods. Epigenetic modifications and miRNA expressions offer new avenues for therapeutic strategies, emphasizing the importance of understanding the cellular mechanisms at play in mare endometrial fibrosis.

The current investigation aimed to explore the effects of liquid extract (MDLE) as the primary component of an extender for breeder rooster semen over different periods at room temperature. Fifteen breeder roosters (40 weeks of age, average body weight of 2.05±0.12) with confirmed fertility were used. Employing a factorial design (3x4), the treatments consisted of semen and two semen extenders (an experimental based on MDLE and a commercial) subjected to four periods at room temperature post-collection (5, 10, 15 and 20 minutes) with four replicates (tubes) each. All variables evaluated in this study yielding significant results (p<0.05). Analyzed individually, the experimental extender based on MDLE exhibited a linear reduction (p<0.05) in motility and vigor results, while it caused an increase in pH values and percentages of sperm defects evaluated. When compared with semen and commercial extender, the efficiency of MDLE as a semen extender was inferior to that observed with the commercial extender and similar to the results observed with semen . Nonetheless, the experimental extender based on MDLE yielded satisfactory results for up to 15 minutes of storage time. In conclusion, MDLE can be considered as an alternative for composing a roosters' semen extender, maintaining sperm characteristics within acceptable limits for up to 15 minutes at room temperature. However, this experimental extender demonstrated lower efficiency than the commercial extender in maintaining the sperm quality at room temperature across all periods tested.

This study evaluated two surgical sterilization techniques in free-ranging female capybaras ( = 21). The first group underwent uterine horn ligature (HL; = 11), while the second was subjected to partial salpingectomy (S; = 10). We assessed total operative time, incision length, the ease of identifying reproductive structures, the adequacy of exposure for surgical performance through flank or midline approaches, and the extent of abdominal viscera manipulation for each method. The HL method emerged as faster, with an average operative time difference of 16 minutes. In the S group, a flank mini-laparotomy over the ovarian topography facilitated easy exposure of the ipsilateral ovary and uterine tube, enabling ligature and partial resection of the uterine tube but not the uterine horn exposure. However, accessing the contralateral uterine tube without a bilateral incision was impractical, thus prolonging the total operative time due to the need for patient repositioning and new antisepsis procedures. Conversely, a post-umbilical approach for the HL method necessitated only one mini-laparotomy incision, offering ample uterine exposure for hysterotomy in pregnant females. Both methods involved minimal abdominal viscera manipulation and resulted in no fatalities or postoperative complications. Although direct comparison is limited by the distinct sterilization techniques and surgical approaches, this study underscores the challenges and surgical access of each method. Our findings endorse the HL technique as an effective contraception method for female capybaras to prevent the birth of seronegative offspring that could amplify sp., the causative agent of Brazilian spotted fever.

This review elucidates the physiological and endocrinological processes intrinsic to puberty and ovulation induction protocols in and beef heifers. Puberty is a complex physiological event involving gonadotropic and metabolic changes that lead to sexual maturity, first ovulation, and regular reproductive cycles, enabling females to reproduce. Exposure to progesterone-based hormonal protocols, with or without additional hormones, can reduce the age at first ovulation and improve sexual maturity through stimuli in the hypothalamus-pituitary-gonadal (HPG) axis and uterine development. However, inducing puberty differs from inducing ovulation, as it does not ensure the heifer will continue cycling or be ready to establish and maintain pregnancy after hormonal exposure. Regardless of the pharmacological basis, studies consistently report that beef heifers that had a corpus luteum (CL) prior to the timed-artificial insemination (TAI) protocol, have greater expression of estrus in response to synchronization and greater pregnancy per AI compared to heifers without a CL. The combination of P4 and E2 significantly impacts uterine development, increasing reproductive efficiency. Exposure to P4 causes a positive effect on inducing ovulation. However, studies indicate that the addition of E2 esters at the time of P4 device removal increases the ovulation rate. In general, the studies showed that fertility varied according to the type of the ovulation induction protocol used, but with inconsistent results. Although ovulation induction protocols are strategic tools to accelerate sexual maturity, a holistic view of the entire system is extremely important, combining integration with genetics and nutrition to enhance the reproductive outcomes of beef heifers. Future research is needed to understand and refine these protocols, driving the efficiency of beef cattle production systems.

Anticancer therapy often leads to premature ovarian insufficiency (POI) and infertility due to the extreme sensitivity of the ovarian follicle reserve to the effects of chemotherapy. Withanolides are known for their cytotoxic effect on cancer cells and low cytotoxicity on non-malignant or healthy cells. Therefore, this study aimed to investigate the effects of three withanolides derivatives: 27-dehydroxy-24,25-epoxywithaferin A (WT1), 27-dehydroxywithaferin A (WT2), and withaferin A (WTA) on fertility, and the ovarian preantral follicles of young female mice. To achieve this, mice received 7 intraperitoneal doses of WT1, WT2, or WTA at a concentration of 2 mg/kg (Experiment I) and 5 or 10 mg/kg (Experiment II) over 15 alternate days. , two days after administration of the last dose, half of the mice were mated to evaluate the effects of withanolides on fertility. The other half of the mice, as well as all mice from , were sacrificed for histological, inflammation, senescence, and immunohistochemical analyses of the follicles present in the ovary. Regardless of the administered withanolide, the concentration of 2 mg/kg did not show toxicity on the follicular morphology, ovarian function, or fertility of the mice. However, at concentrations of 5 and 10 mg/kg, the three derivatives (WT1, WT2, and WTA) increased follicular activation, cell proliferation, and ovarian senescence without affecting inflammatory cells. Furthermore, at a concentration of 10 mg/kg, the three withanolides showed intensified toxic effects, leading to DNA damage as evidenced by the labeling of γH2AX, activated Caspase 3, and TUNEL. We conclude that the cytotoxic effect of the tested withanolide derivatives (WT1, WT2, and WTA) in the concentration of 2 mg/kg did not show toxicity on the ovary. However, in higher concentrations, such as 10 mg/kg, toxic effects are potentiated, causing DNA damage.

produced embryos exhibit lower viability compared to their counterparts. Mammalian preimplantation embryos have the ability to reach the blastocyst stage in diverse culture media, showcasing considerable metabolic adaptability, which complicates the identification of optimal developmental conditions. Despite embryos successfully progressing to the blastocyst stage, adaptation to suboptimal culture environments may jeopardize blastocyst viability, cryotolerance, and implantation potential. Enhancing our capacity to support preimplantation embryonic development requires a deeper understanding of fundamental embryo physiology, including preferred metabolic substrates and pathways utilized by high-quality embryos. Armed with this knowledge, it becomes achievable to optimize culture conditions to support normal, -like embryo physiology, mitigate adaptive stress, and enhance viability. The objective of this review is to summarize the evolution of culture media for bovine embryos, highlighting significant milestones and remaining challenges.

Embryo transfer in cattle is an increasingly important technique for cattle production. Full attainment of the benefits of the technology will depend on overcoming hurdles to optimal performance using embryos produced in vitro. Given its importance, embryo technology research should become a global research priority for animal reproduction science. Among the goals of that research should be developing methods to increase the proportion of oocytes becoming embryos through optimization of in vitro oocyte maturation and in vitro fertilization, producing an embryo competent to establish and maintain pregnancy after transfer, and increasing recipient fertility through selection, management and pharmacological manipulation. The embryo produced in vitro is susceptible to epigenetic reprogramming and methods should be found to minimize deleterious epigenetic change while altering the developmental program of the resultant calf to increase its health and productivity. There are widening opportunities to rethink the technological basis for much of the current practices for production and transfer of embryos because of explosive advances in fields of bioengineering such as microfluidics, three-dimensional printing of cell culture materials, organoid culture, live-cell imaging, and cryopreservation.

This brief review delves into the topic of follicle culture for embryo production, with a particular emphasis on goat models. Specifically, we examine the main findings from LAMOFOPA-Brazil over the last 20 years, highlighting the challenges posed by oxidative stress and epigenetic changes. Our focus is on strategies to improve follicular development and oocyte maturation. Furthermore, we underscore the valuable role of the antioxidant anethole in optimizing the efficacy of follicle culture and improving outcomes in embryo production.

Gene editing technologies have revolutionized the field of livestock breeding, offering unprecedented opportunities to enhance animal welfare, productivity, and sustainability. This paper provides a comprehensive review of recent innovations and applications of gene editing in livestock, exploring the diverse applications of gene editing in livestock breeding, as well as the regulatory and ethical considerations, and the current challenges and prospects of the technology in the industry. Overall, this review underscores the transformative potential of gene editing in livestock breeding and its pivotal role in shaping the future of agriculture and biomedicine.

Due to high annual culling rates, pig farms require a constant income of replacement gilts. Gilts typically reach puberty at nearly six months of age. Puberty may be induced through early boar exposure, therapy with steroid hormones and chorionic gonadotropins, and optimized by identifying biological predictors and risk factors. Old age at the time of the first mating is associated with an increased risk of premature culling, often attributed to reproductive failures and locomotor problems. While female prolifacy has increased substantially during the last few decades, selecting for litter size to optimize lifetime productivity would be more efficient after two parities. Additionally, uterine capacity and the number of functional teats should be considered in selecting future dams. For each female, the cost-effective number of parities at removal is determined by the cumulative number of pigs born and weaned during the total herd days.

The study aimed to evaluate the effectiveness of combining two injectable progesterone (iP4) formulas for estrous synchronization in ewes and to compare it with traditional intravaginal progesterone devices. Additionally, the study assessed whether the inclusion of GnRH enhances the reproductive outcomes of the iP4 treatment. Two experiments were conducted. In the first experiment, 20 Santa Inês ewes were divided into two groups: one group received intravaginal progesterone devices, and the other received combined long-acting and short-acting iP4. In the second experiment, 30 Corriedale ewes were divided into two groups: one received the combined iP4 with GnRH, and the other without GnRH. Estrous, ovulation, follicular populations, and progesterone concentrations were monitored. The combined iP4 treatment induced an artificial luteal phase and produced reproductive responses similar to those obtained with intravaginal devices. In the first experiment, the iP4 treatment tended to result in more synchronized ovulation compared to the control (P=0.095). In the second experiment, adding GnRH enhanced the quality of the corpus luteum, as indicated by increased diameter and vascularization on Day 23 (P=0.047 and P=0.02, respectively). The combined administration of long-acting and short-acting iP4 effectively synchronized estrous in ewes and showed similar efficacy to traditional intravaginal devices. The inclusion of GnRH improved luteal quality, suggesting potential benefits for reproductive management in ewes. Further studies are needed to evaluate the fertility outcomes of these protocols under field conditions.

The cooling of shrimp spermatophores for assisted insemination can enable the transfer of gametes between reproduction laboratories. This study aimed to assess three extenders for cooling spermatophores for assisted insemination. Spermatophores were chilled at 15 °C for 24 or 48 hours using powdered coconut water ACP® (PCW), mineral oil (MO), and sterilized seawater (SSW) as extenders. All treatments demonstrated consistent responses over time. Apparent viability and morphological integrity percentages remained above 60% and 70%, respectively, across treatments and storage durations. Focusing on diluents, normal cell percentages for MO, SSW, and PCW treatments were 74.9±9.20%, 77.3±9.40%, and 78.1±6.35%, respectively, irrespective of storage time. The highest hatching rate was observed in the SSW treatment (80.67±12.01%), which was significantly superior to the PCW treatment (50.15±20.75%). The hatching rates observed in the MO treatment (71.47±18.83%) did not statistically differ from either PCW or SSW treatments. The cooling protocol successfully preserved the spermatophores' ability to maintain favorable levels of apparent viability, normal morphology, and hatching rates after 48 hours of storage at 15 °C using mineral oil, seawater, or ACP® as extenders. Sterilized seawater emerged as the most efficient diluent, delivering superior hatching rates following artificial insemination.

This conference celebrates the 40th anniversary of AETE. Over the past 40 years, AETE has served as a forum for scientists, practitioners, and students working in assisted animal reproduction in livestock species. AETE conferences have reflected developments in the field, from basic to applied science, as well as regulatory changes in assisted animal reproduction practices. Europe has led the way in these developments for many years, progressing from artificial insemination, embryo transfer, and cryopreservation to semen sexing, in vitro production of embryos, cloning by nuclear transfer, genomic selection, and the rescue of highly endangered species. These significant contributions were made possible by the support of funding agencies, both at the national and European levels, promoting cooperation between scientists and practitioners. Assisted reproduction, and animal breeding more generally, face opposition from various groups, including animal rights activists, vegetarians, proponents of organic farming, environmentalists, certain political parties, and increasing regulatory burdens. These challenges seriously affect funding for scientific research, the work of practitioners, and the breeding industry as a whole. It is crucial to invest time and resources in communication to remind the public, politicians, and regulators of the achievements in this field and the contributions made to the food supply chain and the care of the rural and natural environment.

The productivity of the beef and dairy industries depends directly on the reproductive efficiency and genetic gain of the herd, which are directly associated with the appropriate use of Assisted Reproductive Technologies (ARTs). The objective of this review is to show from a Brazilian perspective the evolution over the last 40 years of ARTs related to ovulation resynchronization programs and embryo transfer in cattle. Despite significant improvements and high fertility obtained in timed artificial insemination (TAI) protocols (Sales et al., 2024 - Part I), the improvement of the use of embryos, development of resynchronization programs, and the advance in Doppler ultrasonography (Doppler-US) for reproductive assessments of bovine females were the ARTs that presented the greatest relevance on reproductive effectiveness in cattle. In the last seven years, the embryo transfer (ET) technology using -produced (IVP) embryos took over the conventional ET of produced embryos after donor's superovulation. Also, procedures and pregnancy rates after ET of IVP embryos were improved in dairy and beef operations. The Doppler-US allows the identification of non-pregnant females at an early stage based on the evaluation of blood perfusion of the corpus luteum. Recent studies in beef and dairy cows indicate satisfactory accuracy when Doppler-US is used at 20-22 days after TAI. Consequently, super-early resynchronization programs have been developed and are being implemented in commercial programs, thereby facilitating earlier conception through the use of semen from superior bulls, providing genetic and economic improvements in herds. Likewise, the assessment of luteal function by Doppler-US allows the selection of embryo recipients with greater receptivity, and consequently may increase the effectiveness of timed ET programs.

This study explored the migration of follicular fluid (FF)-derived extracellular vesicles (EVs) of the uterine environment to the bloodstream and their interaction with neutrophils and . For the experiment, six Nellore heifers () received an intrauterine infusion seven days after ovulation with 1X PBS only (sham group; n=1), 1X PBS stained with lipophilic dye PKH26 (control group; n=2), or FF-derived EVs stained with PKH26 (treated group; n=3). Plasma was collected at 0, 10, 30, 60-, 180-, 360-, 720-, and 1440-min post-infusion to obtained EVs for analysis by nano flow cytometry. Labeled EVs were present in the bloodstream at 30- and 60-min post-infusion in the treatment group. Additionally, plasma derived-EVs from all groups were positive for Calcein-AM, Alix, Syntenin, and Calnexin, which confirm the presence of EVs. The second experiment utilized the plasma-derived EVs from the heifers from 30 and 60 min timepoints to evaluate if neutrophils can uptake EVs . As results, it was possible to observe the presence of labeled EVs in neutrophils treated with plasma derived-EVs from the treatment group. In summary, our results suggest that labeled EVs can migrate from the uterine environment rapidly and interact with circulating immune cells in bovine.

plays a crucial role in mammalian spermatogenesis. Here, we integrated single-molecule long-read and short-read sequencing to comprehensively examine expression patterns in adult Baoshan pig (BS) testes. We identified the most important transcript ENSSSCT00000000120 of , obtaining its full-length coding sequence (CDS) spanning 1254 bp. Gene structure analysis located on pig chromosome 5 with 14 exons. Protein structure analysis reflected that PICK1 consisted of 417 amino acids containing two conserved domains, PDZ and BAR_PICK1. Phylogenetic analysis underscored the evolutionary conservation and homology of PICK1 across different mammalian species. Evaluation of protein interaction network, KEGG, and GO pathways implied that interacted with 50 proteins, predominantly involved in glutamatergic synapses, amphetamine addiction, neuroactive ligand-receptor interactions, dopaminergic synapses, and synaptic vesicle recycling, and PICK1 exhibited significant correlation with DLG4 and TBC1D20. Functional annotation identified that PICK1 was involved in 9 GOs, including seven cellular components and two molecular functions. ceRNA network analysis suggested BS was regulated by seven miRNA targets. Moreover, qPCR expression analysis across 15 tissues highlighted that was highly expressed in the bulbourethral gland and testis. Subcellular localization analysis in ST (Swine Tesits) cells demonstrated that PICK1 significantly localized within the cytoplasm. Overall, our findings shed new light on 's role in BS reproduction, providing a foundation for further functional studies of .

Over the past 40 years, assisted reproductive technologies (ARTs) have grown significantly in scale and innovation, from the bovine embryo industry's shift from in vivo derived to in vitro produced embryos and the development of somatic cell-based approaches for embryo production. Domestic animal models have been instrumental in the development of ARTs for wildlife species in support of the One Plan Approach to species conservation that integrates in situ and ex situ population management strategies. While ARTs are not the sole solution to the biodiversity crisis, they can offer opportunities to maintain, and even improve, the genetic composition of the captive and wild gene pools over time. This review focuses on the application of sperm and embryo technologies (artificial insemination and multiple ovulation/in vitro produced embryo transfer, respectively) in wildlife species, highlighting impactful cases in which significant progress or innovation has transpired. One of the key messages following decades of efforts in this field is the importance of collaboration between researchers and practitioners from zoological, academic, governmental, and private sectors.

The aim of the study was to evaluate the use of Acustic Radiation Force Impulse (ARFI) elastography in mammary parenchyma and supramammary lymph nodes, for detection of active mastitis in sheep with naturally infected chronic fibrous lesions. 27 female sheep were included and B-mode ultrasound and ARFI elastography images were obtained, acquiring qualitative (echogenicity and echotexture) and quantitative (shear rate, depth and short/long axis ratio) variables of 48 mammary glands. The glands were divided into three experimental groups: control group (CG) - healthy animals; LSCC- animals that presented fibrous lesions and SCC (somatic cell count) less than 500 x 10 cls/mL; HSCC: animals that presented fibrous lesions and SCC (somatic cell count) more than 500 x 10 cls/mL; The qualitative variables using B-mode ultrasonography, including echotexture and echogenicity, showed no significant differences between the evaluated groups and tissues (p = 0.9336 and p = 0.233, respectively) .In healthy areas of the gland, it was an increasing in shear wave velocity (SWV) in LSCC than in HSCC (p=0.04). When comparing the fibrosis in the LSCC and HSCC groups with their respective normal areas, the velocity increased in both groups: LSCC (p= 0,0007) and HSCC (p= 0,0001). When comparing the areas of fibrosis in LSCC and HSCC with the CG parenchyma, there was an increase in LSCC (p=0.001) and HSCC (p=0.0001). B-mode ultrasound indicate predominance of hypoechoic echogenicity in lymph nodes and reduced short/long axis ratio in cases of active subclinical mastitis. The supramammary lymph node showed increased SWV when comparing the CG with HSCC groups (p=0.02) and GC with LSCC (p=0.04). B-mode ultrasonography is useful for evaluating the mammary parenchyma, however, its application as a standalone diagnostic technique is not recommended. ARFI elastography indicates potential cutoff points for differentiating subclinical mastitis from healed mastitis, highlighting its importance as a tool for distinguishing normal areas from fibrous parenchymal areas. While this study did not establish specific cutoff points due to sample size limitations, further research with larger sample sizes could explore and define these critical thresholds.

The oviduct and uterus provide an optimal environment for early embryo development, where effective communication between the embryo and the maternal reproductive tract is crucial for establishing and maintaining pregnancy. Oviductal and uterine-derived EVs play pivotal roles in this maternal-embryonic communication and in facilitating early embryo development. However, despite the ability of in vitro culture methods to produce viable embryos, the lack of exchange between the embryo and the mother often results in lower-quality embryos than those derived in vivo. Therefore, there is a pressing need to increase our understanding of the physiological mechanisms underlying embryo interaction with the oviduct and endometrium through EVs and to develop models capable of mimicking the in vivo environment. This review aims to provide up-to-date insights into the communication between the mother and pre-implantation bovine embryo, exploring their applications and perspectives in the field.

Oviduct fluid extracellular vesicles (oEV) are essential for periconceptional events. The presence of EV has already been identified in the oviduct fluid (OF) from mammalian species, except in caprine. Therefore, this study aimed to isolate and characterize the caprine oEV (coEV). Initially, in Experiment 1, coEV were isolated from the OF of either each animal individually or from a pool of three animals. In experiment 2, coEV were isolated during the follicular or luteal phases of the estrous cycle. The coEV were characterized by size distribution, polydispersity index (PDI), and zeta potential using dynamic light scattering (DLS) analysis, as well as, by transmission electron microscopy (TEM) and dot blotting (DB). Our results indicated that the physicochemical characteristics of the coEV were similar (P > 0.05), regardless of the isolation method (individual or pool). However, coEV collected during the luteal phase were larger (P < 0.05) than those during the follicular phase. The TEM showed spherical and cup-shaped particles, characteristic of exosomes. The DB revealed the presence of exosomal proteins involved in the biogenesis of coEV. In conclusion, it is possible to isolate and characterize coEV from a single caprine female and the estrous cycle phase influences the vesicles average size and PDI.