The success of artificial insemination (AI) in the swine industry relies on conserving the quality of boar sperm during liquid storage, as boar spermatozoa are prone to oxidative stress due to the high polyunsaturated fatty acid content and lack of antioxidant defenses. Sperm motility, viability, acrosome integrity, and DNA stability are all affected by the increases in reactive oxygen species (ROS) during storage, which lowers fertility. Ethyl pyruvate (EP), a stable derivative of pyruvate, has good antioxidant properties and has been shown to protect sperm quality in vivo. Its effects on boar sperm during in vitro preservation have not yet been investigated. This study investigated the effect of different concentrations of EP (0.1–1 mM) in Beltsville thawing solution at 17°C on the sperm quality parameters of boar spermatozoa over five days. Changes in sperm motility, viability, acrosome integrity, chromatin stability, and ROS were observed. The results showed that boar spermatozoa stored with 0.25–0.75 mM EP showed a significant increase in sperm motility, viability, acrosome integrity, and chromatin stability compared with the control (without EP) and 1 mM EP for 5 days. Compared to the control and 1 mM EP, ROS levels statistically decreased in sperm stored in 0.25–0.75 mM EP on both storage days 3 and 5. Our findings demonstrated that 0.25–0.75 mM of EP could enhance the boar sperm quality and mitigate the oxidative stress during liquid storage, thus revealing a strategy to improve fertility rates during AI in pig production.

Human dermal fibroblasts (HDFs) play a critical role in maintaining skin integrity and promoting tissue repair, but are highly susceptible to apoptosis under stress conditions such as nutrient deprivation. Adipose-derived stem cells (ADSCs) have emerged as a promising therapeutic option due to their regenerative potential and ability to secrete bioactive factors. In this study, we investigated the effect of ADSC-derived paracrine signaling on apoptosis in HDFs and explored the underlying molecular mechanisms. Using a Transwell co-culture system, we found that ADSCs significantly reduced apoptosis in HDFs subjected to low-serum stress, as confirmed by APOPercentage™ staining and the expression of apoptosis-related proteins. Among several soluble factors secreted by ADSCs, hepatocyte growth factor (HGF) exhibited the most pronounced time-dependent increase in culture supernatants. The anti- apoptotic effect of ADSCs was abolished by neutralizing antibodies against HGF, indicating a key role of this factor in mediating fibroblast survival. Further, HDFs were found to express the HGF receptor c-Met at both the mRNA and protein levels. Inhibition of c-Met signaling reversed the cytoprotective effect of ADSCs, suggesting that HGF functions through this receptor. Mechanistically, only the PI3K/AKT pathway—among the major survival pathways tested—was selectively activated in HDFs by ADSC co-culture. Pharmacological inhibition of PI3K/AKT signaling using LY294002 abolished the protective effect, while inhibition of ERK or p38 MAPK had no significant impact. These findings demonstrate that ADSC-derived HGF protects HDFs from stress-induced apoptosis primarily through activation of the c-Met–PI3K/ AKT pathway. This mechanistic insight may provide a basis for the development of stem cell– based therapies aimed at enhancing skin regeneration and fibroblast viability in degenerative or wound-healing contexts.

Targeted protein degradation (TPD) is an emerging therapeutic strategy that leverages the natural protein degradation systems of cells to eliminate disease-associated proteins selectively. Unlike traditional small molecule inhibitors, which merely suppress protein activity, TPD degrades target proteins directly, offering a novel approach to addressing undruggable proteins. The two most extensively studied TPD technologies, proteolysis-targeting chimeras (PROTACs) and molecular glues (MGs), utilize the ubiquitin–proteasome system to induce TPD. PROTACs function as bifunctional molecules that recruit an E3 ubiquitin ligase (E3 ligase) to a target protein, leading to its ubiquitination and subsequent degradation, while MGs enhance protein–protein interactions to facilitate ubiquitination and protein clearance. These approaches have shown promising therapeutic potential in treating cancer, neurodegenerative disorders, and autoimmune diseases, with several compounds currently undergoing clinical trials. Despite these advances, challenges such as limited bioavailability, pharmacokinetic constraints, and target selectivity remain obstacles to the widespread application of TPDbased therapies. Recent developments, including the discovery of novel E3 ligases, linker optimization, and AI-driven drug design, have addressed these limitations, paving the way for the next generation of precision-targeted therapeutics. This paper provides a comprehensive overview of the mechanisms, applications, and future directions of PROTACs and MGs in drug discovery, highlighting their potential to revolutionize modern targeted therapy.

Chronic enteropathy (CE) in dogs presents with persistent symptoms such as vomiting, diarrhea, weight loss, and changes in appetite. Traditional diagnostic methods, including blood tests, ultrasound, and endoscopy, often fail to assess the entire small intestine. Capsule endoscopy offers a non-invasive approach, enabling comprehensive visualization of the gastrointestinal tract and enhancing diagnostic accuracy. In case 1, a 10-year-old Maltese with chronic diarrhea and significant weight loss underwent capsule endoscopy, revealing severe lacteal dilation and moderate gastric erythema, leading to a diagnosis of severe intestinal lymphangiectasia and moderate enteritis. Despite initial corticosteroid therapy and dietary changes, clinical improvement was limited. In case 2, an 11-year-old mixed-breed dog with chronic vomiting, anorexia, and weight loss was assessed. Capsule endoscopy detected moderate gastric erythema, severe small intestinal irregularities, and jejunal bleeding, diagnosing mild esophagitis, moderate gastritis, and severe enteritis with hemorrhage. Treatment with a hypoallergenic diet, corticosteroids, and acid suppressants resulted in gradual symptom relief. This report demonstrates that capsule endoscopy can be utilized as a screening tool for evaluating the mucosal status in patients with CE.

Osteoporosis, characterized by excessive osteoclast activity and disrupted bone remodeling, remains a significant health concern. This study investigated the potential of shiitake mushroom (Lentinula edodes) extracts, prepared with various drying methods, to inhibit osteoclast differentiation and promote bone formation. Mushroom stipe extracts, irrespective of the drying method, effectively suppressed osteoclast differentiation by downregulating nuclear factor of activated T-cells, cytoplasmic 1 expression in osteoclast precursor cells treated with macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-Β ligand (RANKL), whereas the mushroom cap extracts exhibited minimal inhibitory effects. This disparity highlights the importance of part-specific analyses as the stipes contained higher levels of bioactive molecules, including denatonium, which was identified as a key RANKL signaling disrupter. In contrast, the caps lacked the anti-osteoclastogenic activity observed in the stipes, which were notably consistent across the freeze-drying, air-drying, and hot-air drying methods, indicating the stability of the bioactive compounds under various processing conditions. In vivo studies using an osteoporosis-induced zebrafish model further validated the therapeutic potential of the stipe extracts, which restored bone mineralization and mitigated prednisolone-induced bone loss. These findings emphasize the importance of identifying and optimizing specific mushroom parts for their unique therapeutic properties. Shiitake mushroom stipe extracts are a natural product potentially useful in osteoporosis therapy. This therapy would be a promising alternative to conventional treatments, such as bisphosphonates, which are associated with adverse side effects.

A protruding mass was identified in the papilla of the right kidney of a 10-week-old male Sprague-Dawley rat. Microscopically, the neoplastic tissues were consisted of epithelial elements, where basophilic neoplastic cells displayed a high nucleus-to-cytoplasm ratio and formed tubular growth patterns characterized by small, elongated, or convoluted tubules. Blastemal elements were often arranged in aggregates or nests, composed of tightly packed basophilic polygonal to spindloid primitive cells. The surrounding interstitial tissue appeared loose and myxomatous. Based on these histological features, the diagnosis was nephroblastoma. Nephroblastoma is considered as an embryonic tumor originated from metanephric blastemal elements in the renal cortex and typically displays characteristic triphasic patterns. Also, this tumor seldom arises from or remains localized to the renal pelvis. To our literature review, this is the first nephroblastoma occurred at renal papilla in a rat.

A 17-year-old spayed female Shih Tzu dog, weighing 5.0 kg, presented with frequent coughing and respiratory distress. Blood tests revealed mild thrombocytosis, and thoracic ultrasonography and radiography confirmed a significant amount of pleural effusion. However, the thoracic radiographs showed no radiopaque nodules or interstitial patterns indicative of thoracic tumors. Thoracentesis was performed to relieve effusion-induced thoracic pressure, yielding a hemorrhagic serosanguinous pleural fluid. The cytological analysis of this fluid revealed mesothelial cells, supporting the clinical diagnosis of mesothelioma in situ. To address the patient’s clinical symptoms, an aggressive management approach was implemented with chest tube placement to address recurrent pleural effusion after initial thoracentesis. During treatment, the patient exhibited stable health and adapted well to daily life. To the best of our knowledge, this is the first reported case of mesothelioma in situ with hemorrhagic malignant pleural effusion in South Korea. Using a chest tube as an aggressive treatment successfully alleviated dyspnea symptoms and provided symptomatic relief in a patient with mesothelioma in situ.

Hyperoxaluria is a disorder associated with an increased risk of renal stones, one of the most common conditions. For people with hyperoxaluria, there are a limited number of effective therapeutic options. The aim of this study was to examine whether an oxalate-degrading enzyme, oxalate decarboxylase (OxdC), can inhibit crystallization of calcium oxalate (CaOx) in vitro, and whether it can prevent nephrolithiasis caused by CaOx induced by ethylene glycol (EG) in rats. When OxdC was applied at various concentrations to CaOx in vitro, there was a significant reduction in the crystallization of CaOx. The OxdC was found to inhibit crystal formation as well as the formation of crystals that had sharp edges. In animal experiments, rats that had been treated with EG showed impaired renal filtration functions, as well as increased deposition of CaOx crystals and the creation of kidney stones. It has been found that oral administration of OxdC to rats with chronic EG-induced nephrolithiasis that is characterized by CaOx intratubular crystal deposits with hyperoxaluria dramatically reduces the severity of the disease. The results of this study point to a potential therapeutic approach for treating human hyperoxaluria as well as CaOx nephrolithiasis that could be achieved by the oral administration of OxdC.

Following the previous study, which investigated the pharmacological properties of the Technekitty injection (Tc-99m), the toxicity of a single intravenous administration of the Technekitty injection (Tc-99m) and the side effects that may occur at the diagnostic dose were confirmed. The Technekitty injection (Tc-99m) was administered intravenously once at a dose of 0, 0.67, 2.0, and 6.0 mCi/kg to 5 male and female rats per group. Mortality, general symptom observation, and weight measurement were performed for 2 weeks, followed by observation of autopsy findings. There were no deaths, and no statistically significant weight change was observed. No abnormal systemic signs related to the Technekitty injection (Tc-99m) were observed. These results confirmed that Technekitty injection (Tc-99m) can be safely administered intravenously at doses up to 6.0 mCi/kg. Additionally, technetium-99m at an average dose of 2 mCi (74 MBq) has been verified as a diagnostic dose without adverse effects, allowing the Technekitty injection (Tc-99m) to be used safely without side effects at this dosage. This study demonstrates that the Technekitty injection (Tc-99m) has a wide safety margin, supporting its potential for clinical application. Moreover, these findings align with the nonclinical safety standards for radiopharmaceuticals, reinforcing its utility in veterinary medicine. The Technekitty injection (Tc-99m) is expected to be applicable for clinical diagnosis as a veterinary drug in Korea.

Thyroid scanning using technetium-99m (99mTc) is the gold standard for diagnosing feline hyperthyroidism. In cats with an overactive thyroid, a thyroid scan is the most appropriate imaging technique to detect and localize any hyperfunctional adenomatous thyroid tissue. In this study, the pharmacological properties of the Technekitty injection (Tc-99m), developed as a diagnostic agent for feline hyperthyroidism using 99mTc as an active ingredient, were tested in FRTL-5 thyroid follicular cell line and ICR mice. The percentage of cell uptake of the Tc-99m in FRTL-5 thyroid cells was 0.182 ± 0.018%, which was about 6 times higher compared to Clone 9 hepatocytes. This uptake decreased by 38.2% due to competitive inhibition by iodine (sodium iodide). In tissue distribution tests by using ICR mice, the highest distribution was observed in the liver, kidneys, spleen, lungs, and femur at 0.083 hours after administration, and this distribution decreased as the compound was excreted through the kidneys, the primary excretory organ. Maximum distribution was confirmed at 1 hour in the small intestine, 6 hours in the large intestine, and 2 hours in the thyroid gland. Additionally, the total amount excreted through urine and feces over 48 hours (2 days) was 78.80% of the injected dose, with 37.70% (47.84% of the total excretion) excreted through urine and 41.10% (52.16% of the total excretion) through feces. In conclusion, the Tc-99m has the same mechanism of action, potency, absorption, distribution, metabolism, and excretion characteristics as 99mTc used for feline hyperthyroidism in the United States, Europe, and other countries, because the Technekitty injection (Tc-99m) contains 99mTc as its sole active ingredient. Based on these results, the Technekitty injection (Tc-99m) is expected to be safely used in the clinical diagnosis of feline hyperthyroidism.

Non-human primates, due to their high genetic similarity to humans, are used as laboratory animals in biotechnology researches. The growing demand has recently led to a shortage of primate resources, which has become a significant issue both domestically and internationally. This shortage has been further exacerbated by the COVID-19 pandemic. Consequently, the importance of resource conservation through effective primate management is increasing. This requires the establishment of proper quarantine procedures and infectious disease control. Quarantine is an important process that protects not only animal health but also public health significance. Non-human primate quarantine procedures were organized in order. We compared the differences in quarantine procedures not only in Korea but also in various countries such as the US, EU, and Australia. In addition, the etiology, clinical symptoms, diagnosis, and treatment methods of representative infectious diseases of quarantine concern (tuberculosis, monkeypox, monkey immunodeficiency virus, salmonellosis, and shigellosis) were summarized. A literature review of nonhuman primate quarantine procedures in other countries revealed minimal differences in the basic structure. The quarantine periods were similar around 30 days, but we found some differences in details such as legal requirements, documentation forms, and quarantine authorities. These findings are expected to contribute to the development of strategies for improving methods to prevent the spread of infectious diseases and enhancing quarantine management methods.

Myxomatous mitral valve disease (MMVD) in dogs is a heart disease that is characterized by histopathologic changes in cardiomyocytes, which ultimately result in valve degeneration and blood regurgitation due to structural changes in the heart valves. A number of studies have been conducted with the objective of identifying prognostic factors that may influence the prognosis of dogs with MMVD. Nevertheless, there is a paucity of research examining the factors that predict MMVD stage progression as defined by the American College of Veterinary Internal Medicine. The objective of this study was to examine whether there are factors associated with stage progression within one year of diagnosis in dogs diagnosed with subclinical MMVD (stage B1 or B2) using physical examination findings, clinicopathologic biomarkers, and echocardiographic markers. This is a retrospective study of veterinary practice performed at Chungbuk National University Animal Hospital. The electronic medical record of the hospital was searched to obtain clinical records of canine patients diagnosed with subclinical MMVD over an 11-year period. For each patient cohort, a logistic regression analysis was conducted. The variables were initially selected using the backward elimination method, and the optimal logistic regression model was determined by removing the independent variables with the largest variance inflation factor. Among the independent variables examined in this study, heart murmur intensity was identified as a statistically significant predictor of stage progression within one year for subclinical MMVD, a finding that aligns with those of previous studies. No other independent variables were found to be significantly associated with subclinical MMVD stage progression. This is the inaugural exploratory study to concentrate on blood test results, a relatively straightforward and quantifiable test result that can be readily obtained in primary care veterinary clinics, among the factors that may be associated with the progression of subclinical MMVD stages.

Clinical pathology, including hematology and serum chemistry, is an important indicator of biological changes. Animals for inhalation studies are kept in specific chambers and require historical data for accuracy. Age-related characteristics are essential for interpreting experimental results. This study aimed to provide historical clinical pathology data and analyze age-related trends in these parameters. We collected hematological and biochemical parameters from control groups of male and female F344 rats in the 4-, 13-, 26-, and 52-week repeated inhalation toxicity tests. The number of F344 rats from collected control groups were 24, 60, 50, and 25 males and 25, 60, 50, and 25 females in the 4-, 13-, 26-, and 52-week studies, respectively. Mean comparison, correlation analysis and simple linear regression analysis was conducted to reveal age-related trends. Neutrophil count, eosinophil count, neutrophil percentage, monocyte percentage, total protein, albumin, triglyceride, total cholesterol (TCHO) showed increasing trends, whereas lymphocyte count, lymphocyte percent, platelet count, alkaline phosphatase, albumin/globulin ratio, and inorganic phosphate showed decreasing trends in both the mean comparison and regression analyses. TCHO was considered the most affected parameter by aging in both sexes based on statistical results. In this study, we presented clinicopathological data from F344 rats for inhalation toxicity studies. We confirmed aging trends in clinicopathological parameters and identified TCHO as the parameter most affected by aging in F344 rats. These results would be helpful for inhalation research using F344 rats.

We investigated the stress-induced changes in the lipid and hormonal concentrations in plasma, including cytochrome P450 (CYP)-derived oxidative stress in the liver, and the anti-stress effect of Korean Red Ginseng (KRG) water extract in mice. Stress induction using restraint increased the levels of corticosterone (CORT), glucose, total cholesterol (TC), and low-density lipoprotein-cholesterol (LDL-C) while decreasing in the levels of insulin and high-density lipoprotein-cholesterol (HDL-C), compared with those of unstressed mice. Restraint-stress also increased the generation of reactive oxygen species (ROS) in plasma by 5.4-fold. Moreover, the stress resulted in a 2.8-fold higher production of C-reactive protein (CRP) than the control group. In addition, the catalytic activities of CYP1A2 and CYP3A4 in the liver microsomes were stimulated by 5.5- and 3.8-fold, respectively, and concomitant ROS formation was elevated by 4.3-fold in the liver extract, compared to the normal group. In contrast, the KRG treatment (5, 20, or 50 mg/kg/day) to stress-exposed 3 groups alleviated the increased CORT, TC, LDL-C, ROS, and CRP levels and restored the decreased insulin concentrations. The enhanced each ROS in the plasma and liver, and the CYP enzyme activities were also attenuated in KRG-treated mice in a concentration-dependent manner. In conclusion, these results suggest that KRG ameliorates stress-induced detrimental effects on the plasma and liver of treated mice.

Diethylbenzene (DEB) is a colorless flammable liquid composed of a benzene ring and two ethyl substituents. DEBs mostly exist as a mixture of isomers and are mainly used as intermediates and solvents occupationally. Workers may be exposed to DEB inhalation during their occupational activities including manufacturing or processing of materials; however, limited data are available on the risk assessment of DEB mixtures. In this study, male and female Wistar rats were exposed to vapors of a DEB mixture for 13-weeks (6 hr/day, 5 days/ week) at concentrations of 0, 40, 80, and 160 ppm in a whole-body inhalation chamber. Clinical signs, mean body weight, food consumption, bronchoalveolar lavage fluid (BALF), hematology, blood biochemistry, gross findings, organ weights, and microscopic findings were examined to determine the toxicity of DEB mixture. The exposure concentrations in chambers were 39.48 ± 1.13 ppm, 80.43 ± 2.06 ppm, and 160.20 ± 4.42 ppm for the low, medium, and high dose groups, respectively. No changes related to the test substance were observed, including changes in clinical observation, body weight, food consumption, BALF and blood analysis, necropsy findings, absolute and relative organ weights or histopathological analysis. Based on these results, the NOAEC (no-observed-adverse-effect-concentration) of DEB was defined as 160 ppm under the study conditions.

Cell culture is a widely used in vitro tool that enhances our understanding of cell biology, disease mechanisms, drug responses, and the development of tissue engineering. However, there are a number of important drawbacks to conventional two-dimensional (2D) cultures, such as the loss of polarity, altered cell shape, and disruption of cell-extracellular matrix connections. Alternatively, organoids are tissue-engineered, cell-based in vitro models derived from stem cells that can self-organize and differentiate into three-dimensional (3D) structures, recapitulating the morphology and functions of their in vivo counterparts. Bisphenol A (BPA), a ubiquitous industrial chemical, has recently gained recognition as an environmental hazard. Previous research has demonstrated that BPA negatively affects the integrity of the intestinal barrier by triggering programmed cell death and suppressing cell growth in human colonic epithelial cell lines. However, a 2D-based cellular study cannot represent its exposure to multicellular organs. This work investigates the impact of BPA on the structure and function of the intestinal barrier. We examine the effect of BPA on the proliferation and tight junction gene expression with two models: the HT-29 colon cancer cell line and an intestine organoid model and morphological changes of intestinal organoid (I/O). The proliferation was increased in a dose-dependent manner with I/O, but at the same concentration, BPA does not increase the significant number of HT-29 cell respectively. Proliferation-related gene and tight junction gene expression pattern was similar between HT-29 and I/O other than Claudin-4. Therefore, this study offers a more precise depiction of the functional and morphological alterations caused by BPA in comparison to traditional 2D cell cultures.

Inflammatory bowel disease (IBD) is a chronic condition characterized by continuous inflammation of the gastrointestinal tract that varies in intensity over time. IBD is caused by several factors including aberrant gut flora, immunological dysregulation, altered environmental conditions, and genetic variations. However, the pathogenesis of IBD remains unclear. Studies have indicated that an imbalance between T helper 17 (Th17) and regulatory T (Treg) cells contributes significantly to the development of IBD. Intestinal Tregs suppress inflammation and are critical for maintaining tissue homeostasis. Th17 cells are known to play an important role in the development and pathogenesis of IBD and provide non-inflammatory support for the integrity of the intestinal barrier against bacterial and fungal infections. Therefore, the Th17/Treg cell balance is crucial in the pathogenesis of IBD and gut integrity. The microenvironment of the intestinal mucosal immunity is regulated by the secretion of cytokines associated with Th17 cells and Tregs. Several studies have indicated that the gut bacteria contribute to the control of the immune milieu and play a key role in the regulation of Th17 cell development. Intestinal bacteria and cytokines control Th17 cell development. Th17 cells secrete cytokines that regulate the immune microenvironment in the gut mucosa. This review provides an overview of Th17 cells and examines the strategies for treating patients with IBD using Th17 cell-targeted drugs.

This study aimed to investigate the effectiveness of carrageenan (CGN) as an oral immune adjuvant. During the initial research, the inadvertent shallow insertion of an oral gavage needle confirmed CGN’s effect as an adjuvant for esophageal immunization. However, in oral immunization, antibody formation was not observed regardless of CGN’s presence or absence as an adjuvant. Conversely, when bovine serum albumin (used as an antigen) was introduced into the esophagus along with CGN, it resulted in the production of antigen-specific IgG. An exploration was conducted to ascertain whether CGN’s adjuvant effects were associated with prolonging the antigen’s residence time in the esophagus. Upon introducing the antigen into the esophagus without CGN, it was undetectable at two minutes post-introduction. Conversely, when administered with CGN, the antigen remained detectable in the esophagus for up to five minutes post-introduction. To investigate whether this immune response was elicited through mucosal immune mechanisms in the esophagus, the production of IgA, a representative immunoglobulin of mucosal immunity, was assessed. Following esophageal immunization with CGN as an adjuvant, total IgG, IgG1, and IgG2a were detected in serum, while IgA was not detected. These findings suggest that under specific conditions, the esophagus may serve as a site for initiating a novel immune response.

Antimicrobial resistance significantly threatens human and animal health globally, with considerable mortality and economic impact. This study investigated antimicrobial usage in small animal clinics in South Korea, focusing on understanding the trends in prescriptions for therapeutic and preventive purposes. Data were collected from 12 small animal clinics that were analyzed for antimicrobial prescriptions from 2018–2020. A comprehensive dataset was used, including patient signalment, clinical notes, and prescription details, and statistically analyzed using SPSS software. The results indicated that most antimicrobials (93.1%) were prescribed for the treatment of infectious diseases, with a smaller portion (6.9%) used for preventive measures, such as surgery. High prescription rates were observed for the treatment of cutaneous and otological diseases, which may reflect common diseases in companion animals. The study highlighted a higher prescription rate for adult age groups, possibly because of the higher prevalence in those groups. Overall, this study provides valuable insights into common prescription patterns in veterinary practice and underscores the need for more stringent antimicrobial stewardship to curb the rise of antimicrobial resistance. This suggests that ongoing surveillance and education on appropriate antimicrobial use are crucial for optimizing treatment outcomes and minimizing the development of resistance.

The functional roles of plant extracts have been investigated for the treatment of various diseases including subfertility. Recent studies have highlighted the benefits of ashwagandha extract (AE) in enhancing sperm production, boosting testosterone levels, and lowering reactive oxygen species (ROS) levels in mammals. The current study is to examine the effects of the addition of AE to liquid boar semen on sperm quality during storage and its potential application in assisted reproductive technology. A hot water extract of ashwagandha was prepared from the dried powder of ashwagandha roots. Boar spermatozoa were stored in Beltsville thawing solution (BTS) at 17℃ for 5 days, with various concentrations of AE (1–50 mg/mL). During storage, motility, viability, acrosomal integrity and ROS of boar spermatozoa were examined. The results have shown that sperm stored in BTS with varying quantities of AE ranging from 1–20 mg/mL exhibited higher motility compared to those without AE (control) or with 50 mg/mL AE for 5 days. Similarly, sperm viability was better maintained in sperm treated with 1–20 mg/mL AE. Moreover, sperm stored in BTS with AE led to significantly higher acrosomal integrity and chromatin stability rates than sperm stored without AE. Notably, intracellular ROS levels significantly decreased in sperm stored in BTS with AE. Particularly, spermatozoa stored at 10 mg/mL AE exhibited an effective reduction in ROS during storage. These findings suggest the potential role of AE as an additive during sperm storage maintains sperm quality and can be used during subfertility treatment in both animals and humans.