Frontiers in Veterinary Science最新文献

{"title":"Anaplasmosis in the Amazon: diagnostic challenges, persistence, and control of <i>Anaplasma marginale</i> and <i>Anaplasma phagocytophilum</i>.","authors":"Jhorsan David Mauri Pablo, Jakson Jacob Chuquimia Del Solar, Elthon Thomas Hinojosa Enciso, Richard Costa Polveiro, Dielson da Silva Vieira, Eduardo Milton Ramos Sanchez, William Bardales Escalante, Jorge Luis Maicelo Quintana, Rainer Marco Lopez Lapa","doi":"10.3389/fvets.2025.1571694","DOIUrl":"10.3389/fvets.2025.1571694","url":null,"abstract":"<p><p>Anaplasmosis remains a significant threat to livestock production in tropical regions, particularly in the Amazon basin, where ecological complexity and limited veterinary infrastructure challenge effective disease management. This review focuses on <i>Anaplasma marginale</i> and <i>Anaplasma phagocytophilum</i>, the primary species associated with bovine and granulocytic anaplasmosis, respectively. We examine the current state of diagnostic tools, highlighting the limited accessibility of molecular techniques in rural settings and the emerging but underutilized potential of technologies. Persistent infection and antigenic variation are explored as major obstacles for disease eradication and vaccine development. Although live attenuated and inactivated vaccines are in use for <i>A. marginale</i>, none provide sterilizing immunity, and no commercial vaccines exist for <i>A. phagocytophilum</i>. The review evaluates recent advances in recombinant antigens, chimeric constructs, and genetically attenuated strains, as well as future directions involving multiepitope design, novel adjuvants, and next-generation vaccine platforms. Additionally, we assess the role of tick control in disease prevention and emphasize the importance of integrated strategies in regions like the Amazon. Together, these findings underscore the need for context-specific solutions that address the ecological and epidemiological complexity of anaplasmosis in the Amazon basin.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1571694"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12116455/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Pulsatilla chinensis</i> extract alleviate <i>Staphylococcus aureus</i> induced mastitis in mice by regulating the inflammatory response and gut microbiota.","authors":"Yifei Xiang, Ziyang Li, Chengzhi Liu, Zhifei Wei, Xuelian Mo, Yawen Zhong, Ruini He, Zhengmin Liang, Yucheng He, Jiakang He","doi":"10.3389/fvets.2025.1603107","DOIUrl":"10.3389/fvets.2025.1603107","url":null,"abstract":"<p><strong>Introduction: </strong>Subclinical mastitis (SCM) caused by <i>Staphylococcus aureus</i> (<i>S. aureus</i>) is widely prevalent in cattle herds around the world, causing huge losses to the dairy cattle farming industry and dairy product production. Currently, the use of hormones and antibacterial drugs is the most effective treatment method. However, issues such as the increase in drug resistance and residues in dairy products limit their further application. In this study, based on the response surface optimization method, <i>Pulsatilla chinensis</i> extract (PCE) was prepared from <i>Pulsatilla chinensis</i> using ethanol as the medium in a simple, efficient and low-cost way. Its functions were verified both <i>in vitro</i> and <i>in vivo</i>.</p><p><strong>Methods and results: </strong>Through the Oxford cup method, MIC/MBC and co-culture experiments, it was demonstrated that PCE had a good inhibitory effect on the proliferation of four strains of <i>S. aureus</i> <i>in vitro</i>. The <i>in vivo</i> toxicity evaluation proved that PCE had high oral safety. In addition, we screened and established a mastitis model platform for lactating mice to evaluate the expected <i>in vivo</i> effects of PCE. The results showed that pre-treatment with PCE for 7 days significantly reduced the bacterial load and the levels of inflammatory factors (IL-6, IL-1β, TNF-α, MPO) in the mammary gland and blood induced by <i>S. aureus</i>, improved the pathological damage of the mammary gland tissue, and alleviated the occurrence of mastitis in mice by regulating the intestinal microbiota.</p><p><strong>Conclusion: </strong>These results verify that PCE can be used to treat mastitis caused by <i>S. aureus</i>, and thus it is expected to become an excellent alternative to hormones and antibacterial drugs.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1603107"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12116584/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antibacterial and antibiofilm activity of <i>Eucalyptus globulus</i> leaf extract, asiatic acid and ursolic acid against bacteria isolated from bovine mastitis.","authors":"Nicolò Mezzasalma, Costanza Spadini, Chiara Spaggiari, Giannamaria Annunziato, Valentina Andreoli, Alice Prosperi, Lorenzo Mochen, Sandro Cavirani, Stefano Grolli, Simone Taddei, Gabriele Costantino, Clotilde Silvia Cabassi","doi":"10.3389/fvets.2025.1565787","DOIUrl":"10.3389/fvets.2025.1565787","url":null,"abstract":"<p><p>Antibiotics represent the first line therapy for bovine mastitis. However, the increasing prevalence of multidrug-resistant organisms (MDROs) highlights the need for alternative therapeutic approaches. This study evaluated the antimicrobial and antibiofilm activities of <i>Eucalyptus globulus</i> leaf extract (EGL-L), ursolic acid (UA) and asiatic acid (AA) against <i>Staphylococcus aureus</i> (SA), <i>Streptococcus uberis</i> (SU), <i>Streptococcus agalactiae</i> (SAG), and <i>Enterococcus</i> spp. (EN) isolated from bovine mastitis, 39.7% of which were MDROs. The minimal inhibitory concentration (MIC) assay demonstrated that all the compounds exhibited antimicrobial activity against the tested bacteria, including MDROs. However, EGL-L was less effective (<i>p</i> < 0.001) than UA or AA against field strains. UA was more effective against SAG and SU compared to SA (<i>p</i> < 0.001), whereas AA was more effective against SU than SA (<i>p</i> < 0.001). Conversely, EGL-L exhibited similar inhibitory effects on all bacteria. The biofilm-forming ability of the bacterial strains was also assessed, and the minimal biofilm inhibitory concentrations (MBICs) of the compounds were evaluated for moderate and strong biofilm producers. None of the compounds were able to completely inhibit biofilm formation. However, MBIC₈₀ values within the tested concentration range were achieved for 15 out of 32 strains with EGL-L and for 27 out of 32 strains with UA and AA. These findings highlight a promising alternative to conventional antimicrobials for AA and UA, showing potential for topical intramammary use for the control and prevention of bovine mastitis, especially because of their efficacy against biofilm formation. Future research should focus on toxicity assessments and formulation development for potential topical administration.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1565787"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117823/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome editing in mouse spermatogonial stem cell lines targeting the Tex15 gene using CRISPR/Cas9.","authors":"Suheyla Yeşilbostan, Mustafa Yenal Akkurt, Sinan Özkavukçu, Oǧuz Kul, Bengi Çınar","doi":"10.3389/fvets.2025.1599598","DOIUrl":"10.3389/fvets.2025.1599598","url":null,"abstract":"<p><p>CRISPR/Cas9-mediated DNA endonuclease technology has been extensively utilized to introduce targeted genomic mutations for investigating biological processes across various cell types and organisms. In spermatogonial stem cells (SSCs), CRISPR/Cas9 has proven to be an effective tool for elucidating the genetic mechanisms underlying spermatogenesis and infertility. Additionally, it holds potential applications in disease prevention, transgenic animal production, and genetic improvement of livestock. This study aimed to optimize the lipid-based transfection of a lentiviral plasmid vector into SSCs by targeting the Tex15 gene, which is associated with infertility in humans, using CRISPR/Cas9. The efficiency of genome editing was assessed by detecting frameshift indel mutations starting from c.959C in exon 1 of the Tex15 gene using mutation site enzyme cut analysis, sanger sequencing, and <i>in silico</i> analyses. The highest transfection efficiency was achieved with a 1:3.5 DNA:DNAfectin ratio, which was identified as the optimal condition for SSC transfection. CRISPR-Cas9 editing in a monoclonal cell line derived from a single cell yielded high efficiency (model fit <i>R</i> = 0.97). Sequence analysis revealed two possible indel variants, indicating possible heterozygous biallelic editing within the same genome. Our findings demonstrate the potential of SSC-mediated genome editing for generating transgenic animals, enhancing productivity in livestock, and advancing novel therapeutic strategies for genetic disorders in animals and human male infertility.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1599598"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117828/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173760","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Commentary: Oral fluid testing can be used to monitor xenotransplant donor herds for porcine cytomegalovirus/roseolovirus status.","authors":"Joachim Denner","doi":"10.3389/fvets.2025.1571657","DOIUrl":"10.3389/fvets.2025.1571657","url":null,"abstract":"","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1571657"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117819/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173739","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Heterogenous within-herd seroprevalence against epizootic hemorrhagic disease virus type 8 (EHDV-8) after massive virus circulation in cattle in France, 2023.","authors":"Corinne Anthonioz, Yann Abadie, Elodie Reversat, Annie Lafargue, Manon Delalande, Thierry Renaudineau, Laurent Delobel, Nathalie Verdeille, David Ngwa-Mbot, Kristel Gache, Emmanuel Garin, Fabien Corbiere","doi":"10.3389/fvets.2025.1562883","DOIUrl":"10.3389/fvets.2025.1562883","url":null,"abstract":"<p><strong>Background and objectives: </strong>The emergence of Epizootic Hemorrhagic Disease Virus-8 (EHDV-8) in mainland France in 2023 led to thousands of clinical outbreaks in cattle herds and likely led to the natural immunization of a large number of animals. However, uncertainties persist regarding the extent of this immunity, both within herds and across affected regions. This study therefore aimed at investigating the variability of within-herd seroprevalence in clinically affected and non-affected herds across geographical areas with differing levels of disease incidence.</p><p><strong>Methodology: </strong>A study was launched in February 2024 to assess the variability of within-herd seroprevalence in three geographical areas with varying EHDV-8 clinical incidence. A total of 2,763 serums samples from cattle over 24 months in 30 herds with clinical outbreaks and 31 herds without reported clinical case were analyzed using a commercial competitive ELISA.</p><p><strong>Results: </strong>A strong south-north seroprevalence gradient was observed, with the highest animal-level seroprevalence evidenced in the southernmost zone (Pyrenean Piémont) (82.6%, CI 95%: 81.1-83.9), which also experienced the highest incidence of clinical outbreaks. In contrast, significantly lower seroprevalence levels were found in the more northern areas: (zone 2: 11.6, 95% CI: 10.7-12.7; zone 3: 0.3, 95% CI: 0.1-1.1), where clinical outbreaks were less frequent. The within-herd seroprevalence varied widely among herds but was significantly higher in those located in the southernmost zone, compared to other areas. Within each zone, no significant differences in seroprevalence were observed between clinical outbreak herds and non-outbreak herds.</p><p><strong>Discussion: </strong>This study highlights significant geographic and between herd variability in seroprevalence against EHDV-8 after the major virus circulation experienced in 2023, and provides critical insights into regional risks and the potential impact of future EHDV-8 circulation.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1562883"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12116536/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of functional SNP associated with sperm quality in porcine <i>ANXA5</i> that contributes to the growth of immature Sertoli cell.","authors":"Mingyang Han, Diwen Yao, Yuyang Song, Yuhang Liu, Zhihua Chen, Jialian Li, Fenge Li, Xiuqin Yang, Lihe Dai, Buyue Niu","doi":"10.3389/fvets.2025.1576566","DOIUrl":"10.3389/fvets.2025.1576566","url":null,"abstract":"<p><p>AnnexinA5 (ANXA5) has been identified as a positional candidate gene for reproduction and fertility traits in boars, but its role in testicular tissue development, as well as genetic variations remain unclear. The aim of this study was to explore the effect of ANXA5 in the growth of swine Sertoli cells and identify its functional variations. Firstly, the expression of porcine ANXA5 in different tissues was detected by semi-quantitative RT-PCR and its effect on the proliferation of Sertoli cells was evaluated by CCK8, EdU, flow cytometry analyses and qRT-PCR. Then, putative causative variants were screened by integrating in silico analysis and DNA sequencing, and the subsequent association analysis was performed in Largewhite boars. Lastly, dual luciferase reporter assay was used to clarify the effect of specific SNP or ESR1 on ANXA5 transcription. The results showed that ANXA5 expressed in all the detected tissues, promoted proliferation of Sertoli cells by advancing cell cycle progression from the G1 to S phase and encouraging expression of PCNA. Putative causative variants, including two ns-SNPs within the coding region, and three closely linked SNPs in the promoter region were identified. Statistical analysis showed that the frequency of the T allele at g.-676 T > C, A allele at g.-674C > A, and T allele at g.-105G > T were each 0.75, the heterozygotes of Yorkshire boars had greater sperm motility as compared to TT, AA, and TT animals (<i>p</i> < 0.05). Luciferase reporter analysis suggested g.-105G > T and ESR1 modulated ANXA5 transcription. Taken together, this study demonstrated ANXA5 affected swine immature Sertoli cells growth and g.-105G > T was a candidate genetic marker for reproductive trait of boar.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1576566"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12116668/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Case Report: The first record of <i>Eustrongylides</i> sp. infection in the Chinese alligator (<i>Alligator sinensis</i>).","authors":"Yujun Shuai, Yongkang Zhou, Pingsi Yi, Jinhong Zhao","doi":"10.3389/fvets.2025.1579738","DOIUrl":"10.3389/fvets.2025.1579738","url":null,"abstract":"<p><p>Although digestive tract parasites are widely spread in wild or farmed crocodiles worldwide, only limited data are available on <i>Eustrongylides</i> sp. reported in crocodiles. The Chinese alligator (<i>Alligator sinensis</i>) is endemic to the Yangtze River in China, and only a few parasites have been reported to infect the Chinese alligator. In this study, a nematode was collected in the abdominal fascia of a captive deceased Chinese alligator. Cytochrome oxidase I (COI), internal transcribed spacer region (ITS) and partial small subunit DNA segments (18S) sequences were amplified to further confirm the genetic information of the species. The results showed that the nematode was attributed to the genus <i>Eustrongylides</i>. Overall, this is the first report of <i>Eustrongylides</i> sp. infected in the Chinese alligator, expanding the known host range of this nematode and contributing to a better understanding of its life cycle.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1579738"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12116562/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of key genes regulating brown adipose tissue thermogenesis in goat kids (<i>Capra hircus</i>) by using weighted gene co-expression network analysis.","authors":"Minhao Li, Qingjun Zhu, Haili Yang, Yunyi Hu, Le Zhao, Yongju Zhao","doi":"10.3389/fvets.2025.1525437","DOIUrl":"10.3389/fvets.2025.1525437","url":null,"abstract":"<p><p>Brown adipose tissue (BAT) is crucial for the maintenance of body temperature in newborn animals through non-shivering thermogenesis (NST). However, which kind key genes involved in the regulation of BAT thermogenesis and the internal regulation mechanism of heat production in goat BAT were still unclear. In this study, we analyzed the perirenal adipose tissue transcriptome of Dazu black goats from 0, 7, 14, 21 and 28 days after birth using weighted gene co-expression network analysis (WGCNA) to identify key genes involved in the thermogenesis of BAT. Genes were classified into 22 co-expression modules by WGCNA. The turquoise module exhibited high gene expression in D0, with generally lower expression in the later dates. This pattern is consistent with the rapid color, morphological, and thermogenic changes observed in perirenal adipose tissue shortly after birth. GO functional annotation revealed that the genes in the turquoise module were significantly enriched in the mitochondrion, mitochondrial protein-containing complex, cytoplasm, and mitochondrial inner membrane. KEGG pathway enrichment analysis indicated that these genes were predominantly enriched in the signaling pathways of oxidative phosphorylation, thermogenesis, and TCA cycle. By combining the gene co-expression network analysis of the turquoise module genes and the differentially expression genes (DEG) analysis, we identified 5 candidate key genes (<i>ACO2</i>, <i>MRPS27</i>, <i>IMMT</i>, <i>MRPL12</i>, and <i>TUFM</i>) involved in regulation of perirenal adipose tissue thermogenesis. This finding offer candidate genes that in the regulation of BAT thermogenesis and body temperature maintenance in goat kids.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1525437"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12116553/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173763","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tricaine, eugenol and etomidate for repetitive procedural anesthesia in adult zebrafish, <i>Danio rerio</i>: effect on stress and behavior.","authors":"Suneeta Narumanchi, Sanni Perttunen, Pyry Laine, Riikka Kosonen, Päivi Lakkisto, Mika Laine, Ilkka Tikkanen, Jere Paavola","doi":"10.3389/fvets.2025.1562425","DOIUrl":"10.3389/fvets.2025.1562425","url":null,"abstract":"<p><p>Zebrafish has emerged as a popular animal model in biomedical research. Numerous procedures and interventions require occasionally repetitive anesthesia. Tricaine is the most frequently used anesthetic for zebrafish and its efficacy is well established. However, the safety and efficacy of other anesthetics used for zebrafish require further examination, especially regarding repetitive anesthesia. Hence, we compare three anesthetics: tricaine (150 mg/l), eugenol (55 mg/l) and etomidate (4 mg/l) in wildtype adult zebrafish with and without interventions in the form of intraperitoneal injections. Groups of fish receiving the injections are named as (+ injection). We quantify anesthesia induction and recovery times as well as swimming behavior and cortisol levels as indicators of stress. Swimming behavior is quantified with the novel tank method as tank preference and number of turnings. Adult zebrafish are randomly divided into seven groups; tricaine (<i>n</i> = 15), tricaine (+injection; <i>n</i> = 15), eugenol (<i>n</i> = 15), eugenol (+injection; <i>n</i> = 14), etomidate (<i>n</i> = 15), etomidate (+injection; <i>n</i> = 15) and sham (<i>n</i> = 10), and anesthetized until they reach stage 4 anesthesia, daily for 10 days. Following anesthesia induction, injection groups are given daily intraperitoneal injections with 0.9% saline (4 ml/kg) before transfer to a recovery tank to study the effect of a painful procedure (the intraperitoneal injection) during anesthesia on stress. The novel tank method is used for analyzing behavior at day 2 (beginning), day 5/6 (middle) and day 10 (end). Chronic stress is evaluated by whole-body cortisol measurement at the end of the 10-day experiment. Additionally, acute stress is evaluated by whole-body cortisol measurement 30 min after single anesthesia in five groups: tricaine (<i>n</i> = 5), eugenol (<i>n</i> = 5), etomidate (<i>n</i> = 5), sham (<i>n</i> = 5), and untreated controls (<i>n</i> = 5). We find that fish anesthetized with tricaine recover fast (~ 1.5 min) and show normal swimming behavior. Fish anesthetized with eugenol show recovery time (~ 2.5 min) and swimming behavior similar to that of fish receiving tricaine. Fish anesthetized with etomidate have the longest recovery time (~ 5.5 min) and exhibit stressed swimming behavior. Cortisol levels remain at similar levels. Our study supports the use of tricaine as the anesthetic-of-choice for repetitive anesthesia of short duration in zebrafish, followed by eugenol.</p>","PeriodicalId":12772,"journal":{"name":"Frontiers in Veterinary Science","volume":"12 ","pages":"1562425"},"PeriodicalIF":2.6,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117371/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144173099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}