Journal of Animal Science and Biotechnology最新文献

{"title":"Understanding the diversity and roles of the canine gut microbiome.","authors":"Haram Kim, Yeongjae Chae, Jin Ho Cho, Minho Song, Jinok Kwak, Hyunok Doo, Yejin Choi, Juyoun Kang, Hyunjin Yang, Suyoung Lee, Gi Beom Keum, Suphot Wattanaphansak, Sheena Kim, Hyeun Bum Kim","doi":"10.1186/s40104-025-01235-4","DOIUrl":"10.1186/s40104-025-01235-4","url":null,"abstract":"<p><p>The canine gut microbiome plays a vital role in overall health and well-being by regulating various physiological functions, including digestion, immune responses, energy metabolism, and even behavior and temperament. As such, a comprehensive understanding of the diversity and functional roles of the canine gut microbiome is crucial for maintaining optimal health and well-being. In healthy dogs, the gut microbiome typically consists of a diverse array of bacterial phyla, including Firmicutes, Bacteroidetes, Actinobacteria, Fusobacteria, and Proteobacteria. These microbial communities form a complex ecosystem that interacts with the host to support canine health and homeostasis. A well-balanced microbiome, known as eubiosis, represents an optimized microbial composition that enhances host health and metabolic functions. Eubiosis is shaped by interactions between host physiology and environmental factors. However, dysbiosis, a disruption of eubiosis, can contribute to various health issues, such as weight fluctuations, metabolic disorders, and behavioral changes. Maintaining eubiosis in the canine gut microbiome requires customized management strategies that consider both physiological traits and environmental influences. In this review, we explored the structure and function of the canine gut microbiome, with particular emphasis on its role in health and the key factors that influence and support its maintenance.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"16 1","pages":"95"},"PeriodicalIF":6.3,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12228404/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dihydrosanguinarine enhances tryptophan metabolism and intestinal immune function via AhR pathway activation in broilers.","authors":"Yue Su, Miaomiao Wang, Zhiyong Wu, Peng Huang, Jianguo Zeng","doi":"10.1186/s40104-025-01220-x","DOIUrl":"10.1186/s40104-025-01220-x","url":null,"abstract":"<p><strong>Background: </strong>Tryptophan is essential for nutrition, immunity and neural activity, but cannot be synthesized endogenously. Certain natural products influence host health by modulating the gut microbiota to promote the production of tryptophan metabolites. Sanguinarine (SAN) enhances broiler immunity, however, its low bioavailability and underlying mechanisms remain unclear. This study aimed to decode the mechanisms by which sanguinarine enhances intestinal immune function in broilers.</p><p><strong>Methods: </strong>Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed to identify the main metabolites of sanguinarine in the intestine. Subsequently, equal concentrations of sanguinarine and its metabolites were separately added to the diets. The effects of sanguinarine and its metabolites on the intestinal immune function of broiler chickens were evaluated using 16S rRNA gene amplicon sequencing and tryptophan metabolomics approaches.</p><p><strong>Results: </strong>We determined that dihydrosanguinarine (DHSA) is the main metabolite of sanguinarine in the intestine. Both compounds increased average daily gain and reduced feed efficiency, thereby improving growth performance. They also enhanced ileal villus height and the villus-to-crypt (V/C) ratio while decreasing crypt depth and upregulating the mRNA expression of tight junction proteins ZO-1, occludin and claudin-1. Furthermore, both compounds promoted the proliferation of intestinal Lactobacillus species, a tryptophan-metabolizing bacterium, stimulated short-chain fatty acid production, and lowered intestinal pH. They regulated tryptophan metabolism by increasing the diversity and content of indole tryptophan metabolites, activating the aryl hydrocarbon receptor (AhR) pathway, and elevating the mRNA levels of CYP1A1, CYP1B1, SLC3A1, IDO2 and TPH1. Inflammatory cytokines IL-1β and IL-6 were inhibited, while anti-inflammatory cytokines IL-10 and IL-22, serum SIgA concentration, and intestinal MUC2 expression were increased. Notably, DHSA exhibited a more pronounced effect on enhancing immune function compared to SAN.</p><p><strong>Conclusions: </strong>SAN is converted to DHSA in vivo, which increases its bioavailability. DHSA regulates tryptophan metabolism by activating the AhR pathway and modulating immune-related factors through changes in the gut microbiota. Notably, DHSA significantly increases the abundance of Lactobacillus, a key tryptophan-metabolizing bacterium, thereby enhancing intestinal immune function and improving broiler growth performance.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"16 1","pages":"94"},"PeriodicalIF":6.3,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12232056/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent advances in the application of microbiota transplantation in chickens.","authors":"Haoran Zhao, Luke Comer, Muhammad Zeeshan Akram, Matthias Corion, Yang Li, Nadia Everaert","doi":"10.1186/s40104-025-01233-6","DOIUrl":"10.1186/s40104-025-01233-6","url":null,"abstract":"<p><p>Extensive evidence demonstrates that a healthy and well-balanced gut microbiota profoundly influences host nutrient absorption, immunity, and metabolism. Unlike mammals, early microbiota colonization in commercial poultry largely depends on the environment as chicks hatch in incubators under a relatively sterile environment (egg and incubator sterilization) without maternal-offspring interaction. The early gut microbiota remains unsaturated, providing a critical window for modulation and influencing the subsequent microbiota succession, which may have long-term health outcomes. Microbiota transplantation (MT) involves transferring the microbiota from a donor to a recipient to modulate the recipient's microbiota toward a desired state. Successfully applied in human medicine, MT is also gaining attention in poultry production to modulate intestinal health. This review comprehensively explores factors affecting MT, its mechanisms, and its potential applications in chickens, providing insights for further research and commercial use.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"16 1","pages":"91"},"PeriodicalIF":6.3,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12203725/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144509779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of probiotics-derived extracellular vesicles on livestock gut barrier function.","authors":"Yuhan Zhang, Mengzhen Song, Jinping Fan, Xuming Guo, Shiyu Tao","doi":"10.1186/s40104-024-01102-8","DOIUrl":"10.1186/s40104-024-01102-8","url":null,"abstract":"<p><p>Probiotic extracellular vesicles (pEVs) are biologically active nanoparticle structures that can regulate the intestinal tract through direct or indirect mechanisms. They enhance the intestinal barrier function in livestock and poultry and help alleviate intestinal diseases. The specific effects of pEVs depend on their internal functional components, including nucleic acids, proteins, lipids, and other substances. This paper presents a narrative review of the impact of pEVs on the intestinal barrier across various segments of the intestinal tract, exploring their mechanisms of action while highlighting the limitations of current research. Investigating the mechanisms through which probiotics operate via pEVs could deepen our understanding and provide a theoretical foundation for their application in livestock production.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"15 1","pages":"149"},"PeriodicalIF":6.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11542448/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142592374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dietary supplementation with N-acetyl-L-cysteine ameliorates hyperactivated ERK signaling in the endometrium that is linked to poor pregnancy outcomes following ovarian stimulation in pigs.","authors":"Linghua Cheng, Zhicheng Shi, Yuan Yue, Yue Wang, Yusheng Qin, Wei Zhao, Yupei Hu, Qin Li, Min Guo, Lei An, Shumin Wang, Jianhui Tian","doi":"10.1186/s40104-024-01109-1","DOIUrl":"10.1186/s40104-024-01109-1","url":null,"abstract":"<p><strong>Background: </strong>Exogenous gonadotropin-controlled ovarian stimulation is the critical step in animal reproductive management, such as pig, sheep, bovine and other species. It helps synchronize ovulation or stimulate multiple ovulations. However, a number of evidence indicated an unexpected decrease in pregnancy outcomes following ovarian stimulation. This study aimed to explore the underlying mechanism of the pregnancy defect and develop a practical rescue strategy.</p><p><strong>Results: </strong>Compared with those in the control group, gilts that underwent ovarian stimulation showed a decrease in pregnancy rate, farrowing rate, and total number of piglets born. Stimulated gilts also showed an increase in estradiol (E₂) levels. The supraphysiological E₂ level was correlated with the decrease in the number of piglets born. Furthermore, we found that high levels of E₂ impair uterine receptivity, as shown by the overproliferation of endometrial epithelial cells. In vitro mechanistic studies demonstrated that high levels of E₂ hyperactivate FGF-FGFR-ERK signaling cascade in the uterine endometrium, and in turn induces overproliferation of endometrial epithelial cells. Of note, N-acetyl-L-cysteine (NAC) supplementation effectively inhibits ERK hyperphosphorylation and ameliorates endometrial epithelial overproliferation. Importantly, in vivo experiments indicated that dietary NAC supplementation, compared with ovarian stimulation group, improves the uterine receptivity in gilts, and significantly increases the pregnancy rate and total number of piglets born.</p><p><strong>Conclusions: </strong>Ovarian stimulation-induced supraphysiological levels of E₂ impairs uterine receptivity by hyperactivating FGF-FGFR-ERK signaling cascade, thereby reducing pregnancy rate and litter size. Supplementing NAC to a conventional diet for gilts ameliorates hyperactivated ERK signaling and improves uterine receptivity, thus rescuing adverse pregnancy outcomes following ovarian stimulation.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"15 1","pages":"148"},"PeriodicalIF":6.3,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11539329/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142585390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The assembly and activation of the PANoptosome promote porcine granulosa cell programmed cell death during follicular atresia.","authors":"Hao Wu, Yingxue Han, Jikang Liu, Rong Zhao, Shizhen Dai, Yajun Guo, Nan Li, Feng Yang, Shenming Zeng","doi":"10.1186/s40104-024-01107-3","DOIUrl":"10.1186/s40104-024-01107-3","url":null,"abstract":"<p><strong>Background: </strong>Follicular atresia significantly impairs female fertility and hastens reproductive senescence. Apoptosis of granulosa cells is the primary cause of follicular atresia. Pyroptosis and necroptosis, as additional forms of programmed cell death, have been reported in mammalian cells. However, the understanding of pyroptosis and necroptosis pathways in granulosa cells during follicular atresia remains unclear. This study explored the effects of programmed cell death in granulosa cells on follicular atresia and the underlying mechanisms.</p><p><strong>Results: </strong>The results revealed that granulosa cells undergo programmed cell death including apoptosis, pyroptosis, and necroptosis during follicular atresia. For the first time, we identified the formation of a PANoptosome complex in porcine granulosa cells. This complex was initially identified as being composed of ZBP1, RIPK3, and RIPK1, and is recruited through the RHIM domain. Additionally, we demonstrated that caspase-6 is activated and cleaved, interacting with RIPK3 as a component of the PANoptosome. Heat stress may exacerbate the activation of the PANoptosome, leading to programmed cell death in granulosa cells.</p><p><strong>Conclusions: </strong>Our data identified the formation of a PANoptosome complex that promoted programmed cell death in granulosa cells during the process of follicular atresia. These findings provide new insights into the molecular mechanisms underlying follicular atresia.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"15 1","pages":"147"},"PeriodicalIF":6.3,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11536665/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142577370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Natural plant polyphenols contribute to the ecological and healthy swine production.","authors":"Huadi Mei, Yuanfei Li, Shusong Wu, Jianhua He","doi":"10.1186/s40104-024-01096-3","DOIUrl":"10.1186/s40104-024-01096-3","url":null,"abstract":"<p><p>The absence of trace amounts of natural bioactive compounds with important biological activities in traditional dietary models for global farm animals, coupled with an incomplete theoretical system for animal nutrition, has led to unbalanced and inadequate animal nutrition. This deficiency has adversely impacted animal health and the ecological environment, presenting formidable challenges to the advancement of the swine breeding industry in various countries around the world toward high-quality development. Recently, due to the ban of antibiotics for growth promotion in swine diets, botanical active compounds have been extensively investigated as feed additives. Polyphenols represent a broad group of plant secondary metabolites. They are natural, non-toxic, pollution-free, and highly reproducible compounds that have a wide range of physiological functions, such as antioxidant, anti-inflammatory, immunomodulatory, antiviral, antibacterial, and metabolic activities. Accordingly, polyphenols have been widely studied and used as feed additives in swine production. This review summarizes the structural characteristics, classification, current application situation, general properties of polyphenols, and the latest research advances on their use in swine production. Additionally, the research and application bottlenecks and future development of plant polyphenols in the animal feed industry are reviewed and prospected. This review aims to stimulate the in-depth study of natural plant polyphenols and the research and development of related products in order to promote the green, healthy, and high-quality development of swine production, while also providing ideas for the innovation and development in the theoretical system of animal nutrition.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"15 1","pages":"146"},"PeriodicalIF":6.3,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11533317/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142570208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Embryotrophic effect of exogenous protein contained adipose-derived stem cell extracellular vesicles.","authors":"Seonggyu Bang, Ahmad Yar Qamar, Sung Ho Yun, Na-Yeon Gu, Heyyoung Kim, Ayeong Han, Heejae Kang, Hye Sun Park, Seung Ii Kim, Islam M Saadeldin, Sanghoon Lee, Jongki Cho","doi":"10.1186/s40104-024-01106-4","DOIUrl":"10.1186/s40104-024-01106-4","url":null,"abstract":"<p><strong>Background: </strong>Extracellular vesicles (EVs) regulate cell metabolism and various biological processes by delivering specific proteins and nucleic acids to surrounding cells. We aimed to investigate the effects of the cargo contained in EVs derived from adipose-derived stem cells (ASCs) on the porcine embryonic development.</p><p><strong>Methods: </strong>ASCs were isolated from porcine adipose tissue and characterized using ASC-specific markers via flow cytometry. EVs were subsequently extracted from the conditioned media of the established ASCs. These EVs were added to the in vitro culture environment of porcine embryos to observe qualitative improvements in embryonic development. Furthermore, the proteins within the EVs were analyzed to investigate the underlying mechanisms.</p><p><strong>Results: </strong>We observed a higher blastocyst development rate and increased mitochondrial activity in early stage embryos in the ASC-EVs-supplemented group than in the controls (24.8% ± 0.8% vs. 28.6% ± 1.1%, respectively). The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay of blastocysts also revealed significantly reduced apoptotic cells in the ASC-EVs-supplemented group. Furthermore, through proteomics, we detected the proteins in ASC-EVs and blastocysts from each treatment group. This analysis revealed a higher fraction of proteins in the ASC-EVs-supplemented group than in the controls (1,547 vs. 1,495, respectively). Gene analysis confirmed that ASC-EVs showed a high expression of tyrosine-protein kinase (SRC), whereas ASC-EVs supplemented blastocysts showed a higher expression of Cyclin-dependent kinase 1 (CDK1). SRC is postulated to activate protein kinase B (AKT), which inhibits the forkhead box O signaling pathway and activates CDK1. Subsequently, CDK1 activation influences the cell cycle, thereby affecting in vitro embryonic development.</p><p><strong>Conclusion: </strong>ASC-EVs promote mitochondrial activity, which is crucial for the early development of blastocysts and vital in the downregulation of apoptosis. Additionally, ASC-EVs supply SRC to porcine blastocysts, thereby elongating the cell cycle.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"15 1","pages":"145"},"PeriodicalIF":6.3,"publicationDate":"2024-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11531693/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142565434","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects and interactions of dietary lysine and apparent nitrogen corrected metabolizable energy on yellow-feathered broiler breeder hens.","authors":"Yibing Wang, Qin Wang, Xiangtian Yao, Zhongyong Gou, Xiajing Lin, Qili Luo, Shouqun Jiang","doi":"10.1186/s40104-024-01095-4","DOIUrl":"10.1186/s40104-024-01095-4","url":null,"abstract":"<p><strong>Background: </strong>Dietary lysine and apparent nitrogen-corrected metabolizable energy (AMEn) are two key variables affecting the production of breeder hens. In this study, the effects and interactions of lysine and AMEn on yellow-feathered broiler breeder hens were investigated. A total of 720 30-week-old breeder hens were fed in a 5 (lysine: 0.56%, 0.68%, 0.80%, 0.92%, and 1.04%) × 2 (AMEn: 11.50 and 11.10 MJ/kg) factorial arrangement for 12 weeks. The productive performance, reproductive traits, biochemical variables of breeder hens, the amino acid concentration and quality of eggs, and the growth performance of offspring broilers were determined.</p><p><strong>Result: </strong>(1) Dietary lysine had quadratic effects (P < 0.05) on laying rate, average daily egg mass and feed intake/egg mass of breeder hens; birds with 11.50 MJ/kg AMEn (high AMEn) had higher (P < 0.05) BW than those with 11.10 MJ/kg AMEn (low AMEn); (2) dietary lysine significantly affected on the relative ovarian weight (quadratic and linear), and numbers of large yellow follicles (LYF, quadratic); birds with high AMEn had longer fallopian tube and more LYF than those with low AMEn (P < 0.05); (3) dietary lysine had significant effects (linear and quadratic) on eggshell thickness and shell strength of eggs from breeder hens; birds with high AMEn had thinner eggshells and deeper yolk color than those with low AMEn (P < 0.05); (4) there were higher (P < 0.05) contents of protein and concentrations of all measured animo acids (AAs) in eggs from birds fed low AMEn; (5) supplementation with high AMEn to breeder hens significantly increased the hatchability of fertilized eggs; (6) neither dietary lysine level or AMEn affected growth performance of offspring broilers; (7) both dietary lysine level and AMEn significantly affected gonadotropin concentrations and biochemical variables of breeder hens.</p><p><strong>Conclusions: </strong>Dietary lysine had significant influences on productive performance, reproductive traits, and egg quality of yellow-feathered breeder hens. Based on productive performance, the optimal levels of dietary lysine were 0.81% to 0.83%, while 0.71% to 72% lysine was enough to obtain the best quality of breeding eggs. High AMEn was more beneficial to breeder hens for reproductive traits and hatchability of the fertilized eggs, while it showed detrimental effects on eggshell thickness and AA concentrations of breeding eggs.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"15 1","pages":"143"},"PeriodicalIF":6.3,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11529154/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142559582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryopreservation of bovine sperm causes single-strand DNA breaks that are localized in the toroidal regions of chromatin.","authors":"Jordi Ribas-Maynou, Rodrigo Muiño, Carolina Tamargo, Marc Yeste","doi":"10.1186/s40104-024-01099-0","DOIUrl":"10.1186/s40104-024-01099-0","url":null,"abstract":"<p><strong>Background: </strong>Sperm cryopreservation is widely used in the cattle industry, as it allows for disassociating the localization of sires and the collection of semen from the timing of artificial insemination. While freeze-thawing is known to impair sperm DNA integrity, whether the damage induced consists of single- (SSB) or double-strand breaks (DSB) has not been determined. In addition, no previous study has addressed if DNA breaks preferentially reside in specific genome regions such as those forming the toroid linker regions, or are rather spread throughout the regions linked to protamines. The main aim of the present work, therefore, was to elucidate the type and localization of the DNA damage generated by cryopreservation and to evaluate its impact on artificial insemination outcomes in cattle.</p><p><strong>Results: </strong>The incidence of SSB and DSB was evaluated in 12 ejaculates before and after cryopreservation with the Comet assay, and the localization of the DNA breaks was assessed using pulsed-field gel electrophoresis (PFGE). Before cryopreservation, the incidence of SSB was 10.99% ± 4.62% and involved 20.56% ± 3.04% of sperm cells, whereas these figures significantly (P < 0.0001) increased up to 34.11% ± 3.48% and 53.36% ± 11.00% in frozen-thawed sperm. In contrast, no significant differences in the incidence of DSB were observed (P > 0.990) before and after cryopreservation (before: incidence of 13.91% ± 1.75% of sperm DNA affecting 56.04% ± 12.49% of sperm cells; after: incidence of 13.55% ± 1.55% of sperm DNA involving 53.36% ± 11.00% of sperm cells). Moreover, PFGE revealed that the percentage of sperm DNA fragments whose length was shorter than a toroid (< 31.5 kb) was greater (P < 0.0001) after (27.00% ± 4.26%) than before freeze-thawing (15.57% ± 4.53%). These differences indicated that the DNA breaks induced by cryopreservation affect the regions condensed in protamines, which are structured in toroids. On the other hand, in vivo fertility rates were associated to the incidence of SSB and DSB in frozen-thawed sperm (P = 0.032 and P = 0.005), but not with the size of the DNA fragments resulting from these breaks (P > 0.05).</p><p><strong>Conclusion: </strong>Cryopreservation of bovine sperm generates single-strand DNA breaks, which are mainly located in protamine-condensed toroidal regions. The incidence of DNA breaks in cryopreserved sperm has an impact on cattle fertility, regardless of the size of generated fragments.</p>","PeriodicalId":64067,"journal":{"name":"Journal of Animal Science and Biotechnology","volume":"15 1","pages":"140"},"PeriodicalIF":6.3,"publicationDate":"2024-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11470689/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142407269","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}