Shreeya Sharma, Mohammadali Alizadeh, Scott Pratt, Alexis Stamatikos, Khaled Abdelaziz
{"title":"Differential Expression of Key Immune Markers in the Intestinal Tract of Developing Chick Embryos.","authors":"Shreeya Sharma, Mohammadali Alizadeh, Scott Pratt, Alexis Stamatikos, Khaled Abdelaziz","doi":"10.3390/vetsci12020186","DOIUrl":"https://doi.org/10.3390/vetsci12020186","url":null,"abstract":"<p><p>Research on the immunological development of lymphoid organs in chicks has been extensive, yet a significant gap exists in our understanding of innate immunity during embryonic life within the intestinal tract. This study investigated the developmental trajectory of intestinal immunity in chick embryos by evaluating basal gene expression levels of key immune markers at embryonic days (ED) 14, 17, and 20. The results indicated variable expression levels of cytokines, antimicrobial peptides (AMPs), and Toll-like receptor (TLRs) genes throughout the intestinal tract. Most cytokines and chemokines exhibited elevated expression in the cecum, while AMPs, including avian-β-defensins (AvBDs) and cathelicidins (CATHs) genes, showed increased levels in the jejunum at ED20. The findings from the developmental trajectory analysis of these genes revealed elevated expression levels of cytokines, including interferon (IFN)-γ, interleukin (IL)-6, IL-13, and transforming-growth factor (TGF)-β in the cecum at ED20. However, no consistent patterns were observed for AvBDs, CATHs, and TLRs, as their expression varied across different developmental stages of the chick embryo. These findings significantly contribute to our understanding of intestinal immune system development in chick embryos and provide a foundation for further research aimed at enhancing immune capabilities, especially in segments with lower expression levels of immunomodulatory genes.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Geovanni Dantas Cassali, Karen Yumi Ribeiro Nakagaki, Marisa Salvi, Marina Possa Dos Reys, Marcos André Nino Rocha, Cecilia Bonolo de Campos, Enio Ferreira, Angelica Cavalheiro Bertagnolli Rodrigues, Diego Carlos Dos Reis, Karine Araujo Damasceno, Alessandra Estrela-Lima
{"title":"Canine, Feline, and Murine Mammary Tumors as a Model for Translational Research in Breast Cancer.","authors":"Geovanni Dantas Cassali, Karen Yumi Ribeiro Nakagaki, Marisa Salvi, Marina Possa Dos Reys, Marcos André Nino Rocha, Cecilia Bonolo de Campos, Enio Ferreira, Angelica Cavalheiro Bertagnolli Rodrigues, Diego Carlos Dos Reis, Karine Araujo Damasceno, Alessandra Estrela-Lima","doi":"10.3390/vetsci12020189","DOIUrl":"https://doi.org/10.3390/vetsci12020189","url":null,"abstract":"<p><p>In veterinary medicine, mammary tumors are the most common neoplasms in female dogs and the third most frequent in cats, representing a significant challenge. Efforts have been directed toward adopting standardized diagnostic criteria to better understand tumor behavior and progression in these species. Meanwhile, the use of animal models has substantially advanced the understanding of comparative mammary carcinogenesis. These models provide critical insights into factors responsible for the disease in humans, with the expectation that such factors can be identified and controlled. In this context, this review presents a work based mainly on articles published by a research group specializing in mammary pathology (Laboratory of Comparative Pathology-Department of General Pathology-ICB/UFMG) and its collaborators, complementing their results with literature findings. The publications were categorized into animal research, experimental research, and human research. These studies addressed topics such as diagnosis, prognostic and predictive factors, tumor microenvironment, inflammation associated with tumors, treatment approaches, and factors influencing tumor growth. The conceptual network analysis underscores the importance of in vivo breast cancer models, both experimental and spontaneous, for understanding tumor progression mechanisms and therapeutic responses, offering valuable contributions to veterinary and human oncology.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sophia Ballard, Alexandria Montgomery, Ian Rose, Shawn Lockhart, Emilio DeBess, Luiz E Bermudez
{"title":"Epidemiological Study of <i>Cryptococcus gattii</i> Complex Infection in Domestic and Wild Animals in Oregon.","authors":"Sophia Ballard, Alexandria Montgomery, Ian Rose, Shawn Lockhart, Emilio DeBess, Luiz E Bermudez","doi":"10.3390/vetsci12020185","DOIUrl":"https://doi.org/10.3390/vetsci12020185","url":null,"abstract":"<p><p>The members of the <i>Cryptococcus gattii</i> species complex are the etiologic agents of potentially fatal human infection. <i>C. gattii</i> causes disease in both immunocompetent and immunocompromised hosts. In the early 2000s, infection caused by <i>C. gattii</i> emerged in the Pacific Northwest of the US. While many studies have been published about the human infection, the epidemiological characteristics of the infection in animals, with a possible role in human infection, have not been in investigated. Cases of <i>C. gattii</i> diagnosed in animals in Oregon from 2008 to 2019 were cataloged by county, species of animal, site of the infection, season of the year, and <i>C. gattii</i> genotype. One hundred and nine cases were diagnosed, and among the genotypes of <i>C. gattii,</i> VGII (<i>Cryptococcus deuterogatti</i>) with the genotypes VGIIa, VGIIb, and VGIIc was responsible for 98% of the cases. VGIIa was identified in more than 50% of the animals, and <i>Cryptococcus bacilliporus</i> (VGIII) was only isolated from cat patients. The majority of the infections were diagnosed in dogs and cats, although caprines, equines, camelids, ovines, and elk were also seen with the disease. The most common site of infection in dogs was the brain; that in cats was the nasal cavity and the skin, while the lung was the most affected site in caprines, equines, camelids and elk. Marion and Lane Counties account for the majority of the infections, followed by Clackamas, Benton, and Multnomah Counties. The infection was predominantly identified during the Fall and Winter months, except for Benton County, where it was seen more commonly during the Summer months. This study reviews all the cases identified by the Department of Public Health and by the veterinarians in Oregon in the years between 2008 and 2019.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ahsa Oh, Sung-Min Nam, Sang-Soep Nahm, Ki-Dong Eom, Woosuk Kim
{"title":"Comparison of Medical Imaging Quality Related to Embalming Solutions in Canine Cadavers.","authors":"Ahsa Oh, Sung-Min Nam, Sang-Soep Nahm, Ki-Dong Eom, Woosuk Kim","doi":"10.3390/vetsci12020188","DOIUrl":"https://doi.org/10.3390/vetsci12020188","url":null,"abstract":"<p><p>(1) Background: This study evaluated the quality of medical imaging acquired using radiography, ultrasonography, and computed tomography (CT) in canine cadavers prepared with formalin solution, Thiel solution (TS), and saturated salt solution (SS). Additionally, the duration whereby image quality could be properly reproduced was investigated. (2) Methods: Nine dog cadavers, divided into three groups per embalming solution, were imaged pre- and post-embalming at 6 and 12 weeks using three different imaging modalities. Image quality was qualitatively scored based on human medicine guidelines criteria. The score was assigned as follows: \"0 = not invisible, 1 = poorly invisible, 2 = adequately reproduced, 3 = very well reproduced\". (3) Results: On plain radiography, most scores were > 1.34, although the quality of all embalming solutions in thoracic and abdominal images decreased over time. Using ultrasound to image the liver, spleen, and kidney, the SS group was evaluated as poorly visible and scored < 0.82 at 6 and 12 weeks. CT images showed a tendency to degrade over time, but all solution groups had properly implemented images and scored > 1.62. (4) Conclusions: Regarding image quality and ease of application, TS is considered the most suitable embalming solution for medical imaging acquisition and is usable over time in all imaging modalities.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504553","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Relationships Among In-Line Milk Fat-to-Protein Ratio, Metabolic Profile, and Inflammatory Biomarkers During Early Stage of Lactation in Dairy Cows.","authors":"Karina Džermeikaitė, Justina Krištolaitytė, Neringa Sutkevičienė, Toma Vilkonienė, Gintarė Vaičiulienė, Audronė Rekešiūtė, Akvilė Girdauskaitė, Samanta Arlauskaitė, Árpád Csaba Bajcsy, Ramūnas Antanaitis","doi":"10.3390/vetsci12020187","DOIUrl":"https://doi.org/10.3390/vetsci12020187","url":null,"abstract":"<p><p>The early lactation phase in dairy cows is characterised by significant metabolic and inflammatory changes. This study aimed to evaluate the relationship between serum nonesterified fatty acids (NEFAs), a marker of negative energy balance (NEB), and serum amyloid A (SAA), an indicator of systemic inflammation. Blood samples were collected from 71 Holstein cows during the transition period 17 (±3) DIM, and serum concentrations of NEFAs and SAA were measured. The results revealed a significant negative correlation between NEFAs and SAA (r = -0.441, <i>p</i> < 0.001), suggesting that increased fat mobilisation may suppress the inflammatory response, thereby increasing the susceptibility to metabolic and infectious diseases. The emerging research indicates a negative association between SAA levels and milk fat-to-protein ratio in dairy cows, particularly under inflammatory conditions. The research indicates that elevated levels of SAA, which is an inflammatory biomarker, are frequently associated with alterations in milk composition, including a reduced fat-to-protein ratio. This study examined the correlations among serum NEFAs, SAA, milk composition, and dairy cattle health. A strong positive correlation was identified between serum NEFAs and milk fat content (r = 0.459, <i>p</i> < 0.001), as well as between serum NEFAs and the milk fat-to-protein ratio (r = 0.516, <i>p</i> < 0.001). Cows with elevated serum NEFA levels (classified as II-NEFA) exhibited significantly higher milk fat content (4.20%) and milk fat-to-protein ratios (1.33) compared to cows with lower serum NEFA levels (I-NEFA class; 3.81% and 1.17, respectively). The data indicate that elevated serum NEFA levels are associated with an increased milk fat synthesis, likely driven by enhanced fat mobilisation during NEB. A significant negative correlation was observed between SAA and both milk fat content (r = -0.426, <i>p</i> < 0.001) and the milk fat-to-protein ratio (r = -0.535, <i>p</i> < 0.001), indicating that inflammation may impair milk fat production. Elevated SAA levels were also associated with increased cow activity (r = 0.382, <i>p</i> < 0.001), suggesting that inflammation may lead to behavioural changes driven by discomfort. Our findings suggest that milk composition reflects the metabolic and inflammatory status of dairy cows and could serve as a non-invasive alternative to blood sampling for assessing energy balance and health. NEB, which typifies early lactation, promotes fat mobilisation, resulting in elevated serum NEFA levels and an increased risk of metabolic disorders such as fatty liver syndrome and ketosis. Moreover, high serum NEFA levels adversely affect immune function, increasing vulnerability to infections such as mastitis. Monitoring milk composition may enable the early detection of NEB and inflammatory conditions, thereby supporting proactive health management. However, further research is necessary to elucidate the role of NEFAs and inflammation in ","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The Occurrence and Meta-Analysis of Investigations on Intestinal Parasitic Infections Among Captive Wild Mammals in Mainland China.","authors":"Xueping Zhang, Hongyu Zhou, Lina Ye, Jiayu Shi, Huiling Zhang, Tangjie Zhang","doi":"10.3390/vetsci12020182","DOIUrl":"https://doi.org/10.3390/vetsci12020182","url":null,"abstract":"<p><p>This study investigated the infection rates of intestinal parasites in captive wild animals from 2000 to 2024 and analyzed the associated risk factors using a meta-analysis. We retrieved 29 studies, with a total of 8421 captive wild mammals, published between 2000 and 2024. We used the DerSimonian-Laird model and calculated infection rate estimates with the variance stabilizing double arcsine transformation. The results revealed that the overall prevalence of gastrointestinal parasitic infections in captive wild mammals in mainland China was found to be 53.9%. The highest infection rate was observed for nematodes at 45.1%. Seasonal subgroup analysis revealed the highest incidence in summer at 61.8% and 61.6% in winter. In the class order Mammalia, the highest infection rate was found in the Primates at 66.5% and similarly high values were determined in several other orders like Artiodactyla (59%), Rodentia (57.1%), Carnivora (53.3%) and extremely low in Proboscidea (19.9%). The highest overall infection rate was recorded in summer. The infection rate of gastrointestinal parasites in captive wild mammals in mainland China is notably high. Additionally, parasitic infection rates in captive animals were found to be lower in economically developed regions of mainland China.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Isolation, Genomic Characterization, and Immunogenicity Evaluation of a G9P[23] Porcine Rotavirus Strain.","authors":"Zixuan Wang, Wen Huang, Gengxuan Yan, Yuan Tian, Chune Wang, Xue Mao, Meng Sun, Lu Zhou, Chong Yu, Haihua Xia","doi":"10.3390/vetsci12020180","DOIUrl":"https://doi.org/10.3390/vetsci12020180","url":null,"abstract":"<p><p>(1) Background: Rotavirus (RV) is the primary pathogen causing diarrhea in infants and young children. The G9-type is an emerging genotype; however, its isolation is challenging, and its immunogenicity to piglets is poorly understood. This study aimed to isolate, characterize, and evaluate the immunogenicity of a PoRVA strain, RHeN2, obtained from the diarrhea samples of piglets from a pig farm in Henan Province, China. (2) Methods: Porcine rotavirus A (PoRVA) was isolated from diarrheal samples of piglets on a farm in Henan Province, China, through sequential passaging on MA-104 cells. Its immunogenicity and cross-neutralization potential were evaluated. Inactivated vaccines were prepared using two agents (formaldehyde and binary ethyleneimine, BEI) and administered intramuscularly to 28-day-old piglets. Dulbecco's Modified Eagle Medium (DMEM) served as the control. (3) Results: A PoRVA strain, G9P[23] type, was successfully isolated and named RHeN2. Full-genome Sanger sequencing revealed the genetic constellation of this strain as G9-P[23]-I5-R1-C1- M1-A8-N1-T1-E1-H1. In addition, the titers of neutralizing antibodies peaked at 45 days post-vaccination, and the immunogenicity of the BEI-inactivated group of vaccines was better than that of the formaldehyde-inactivated group. The RHeN2 (G9P[23]) strain demonstrated a strong cross-neutralization ability against the same G9P[23] and G4P[23] strains and a stronger cross-neutralizing ability against the P[23] strains than against the P[7] strains. (4) Conclusion: The newly isolated G9P[23] strain demonstrates favorable immunogenicity, holding potential as a G9-type vaccine candidate.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Utilizing NF-κB Signaling in Porcine Epithelial Cells to Identify a Plant-Based Additive for the Development of a Porcine Epidemic Diarrhea Virus Vaccine.","authors":"Nguyen-Thanh Hoa, Haroon Afzal, Thu-Dung Doan, Asad Murtaza, Chia-Hung Yen, Yao-Chi Chung","doi":"10.3390/vetsci12020181","DOIUrl":"https://doi.org/10.3390/vetsci12020181","url":null,"abstract":"<p><p>The nuclear factor-kappa B (NF-κB) signaling pathway plays a crucial role in regulating immune responses in epithelial cells. In this study, we established a porcine epithelial NF-κB reporter cell line (PK15-KBR) as an in vitro platform to screen plant-based extracts for their potential use as vaccine adjuvants against porcine epidemic diarrhea virus (PEDV). The NF-κB inducers were further tested for toxicity assessments, either using CCK-8 assays or intramuscular injection in mice, finally followed by vaccination studies to evaluate their adjuvancy. Initial experiments confirmed that TNF-α effectively activated NF-κB signaling in PK15-KBR cells in a dose-dependent manner, validating the platform's reliability at Z' value of 0.68. Of the 224 testers, 3 candidates, including chamomile, mulberry and <i>Boerhaavia diffusa</i>, showed induction activity; however, only chamomile induced a dose-dependent response in PK15-KBR cells. As a proof of concept, chamomile, used as an adjuvant in oral vaccination, demonstrated significantly higher IgG levels at an early stage (day 14, <i>p</i> < 0.05) and enhanced IgA titers. These findings highlight the use of the PK15-KBR cell line in identifying mucosal adjuvants and position chamomile extract as a promising candidate for enhancing vaccine-induced immunity.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143503802","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Marine Inglebert, Martina Dettwiler, Chang He, Enni Markkanen, Lennart Opitz, Arunasalam Naguleswaran, Sven Rottenberg
{"title":"Individualized Pooled CRISPR/Cas9 Screenings Identify CDK2 as a Druggable Vulnerability in a Canine Mammary Carcinoma Patient.","authors":"Marine Inglebert, Martina Dettwiler, Chang He, Enni Markkanen, Lennart Opitz, Arunasalam Naguleswaran, Sven Rottenberg","doi":"10.3390/vetsci12020183","DOIUrl":"https://doi.org/10.3390/vetsci12020183","url":null,"abstract":"<p><p>High-throughput omics approaches have long been used to uncover potential vulnerabilities in human personalized oncology but are often limited by the lack of functional validation. Therefore, we placed our emphasis on functional drug testing using patient-derived organoids (PDOs). However, PDOs generated from tumors mostly lack comparison with matching normal tissue, and the number of testable drugs is limited. Here, we demonstrate how matching the neoplastic and non-neoplastic mammary PDOs derived from the same dog can utilize targeted CRISPR/Cas9 screens to unveil cancer cell specific vulnerabilities. We performed two independent CRISPR/Cas9 dropout screens using sub-libraries targeting the epigenome (<i>n</i> = 1269) or druggable genes (<i>n</i> = 834) in paired PDOs derived from both carcinoma and normal mammary tissues from the same dog. A comparison of essential genes for tumor cells survival identified CDK2 as a functional vulnerability in canine mammary tumors (CMTs) that can be targeted with the PF3600 inhibitor. Additional potential targets were also uncovered, providing insights for personalized cancer treatments in dogs.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Matheus Vicente Silva, Lays Oliveira Rocha, Bruno Augusto Nassif Travençolo, Ednaldo Carvalho Guimarães, Marcelo Emilio Beletti
{"title":"Methods Used to Assess Bull Sperm Chromatin Integrity and Its Correlation with In Vitro Embryo Production Efficiency.","authors":"Matheus Vicente Silva, Lays Oliveira Rocha, Bruno Augusto Nassif Travençolo, Ednaldo Carvalho Guimarães, Marcelo Emilio Beletti","doi":"10.3390/vetsci12020184","DOIUrl":"https://doi.org/10.3390/vetsci12020184","url":null,"abstract":"<p><p>Sperm chromatin analysis is a crucial tool for investigating fertility in bulls, both in the field and in in vitro fertilization. This study aimed to identify efficient, low-cost, and easy-to-apply methods for detecting chromatin integrity alterations. Frozen semen samples from four bulls with varying results in in vitro embryo production (IVEP) were used. The sperm chromatin from these samples was evaluated using different methods: differences in the staining intensity with toluidine blue and the Feulgen reaction (FR), assessed both visually and computationally; visual evaluation of smears stained with acridine orange (AO) using epifluorescence and confocal microscopy; and evaluation using transmission electron microscopy (TEM) of sections positively stained with uranyl and lead. The same samples were utilized for IVEP. The results obtained using each method were correlated with one another and with the cleavage and embryonic development rates achieved in IVEPs. The odds ratio was employed to compare the chromatin alteration identification rates of the studied methods. Overall, chromatin integrity alterations identified using the AO and TEM methods exhibited correlations with the cleavage (r = -1.00) and embryonic development rates (r = -1.00), respectively. Among the methods tested, TEM and AO were the most effective for identifying chromatin integrity alterations that interfere with IVEP efficiency.</p>","PeriodicalId":23694,"journal":{"name":"Veterinary Sciences","volume":"12 2","pages":""},"PeriodicalIF":2.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504516","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}