Clinical, haematological, and biochemical responses of rectal oxygen-ozone administration in sheep

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SOUZA, RENAN S. CARVALHO, CLARISSE S. COELHO, ÂNGELA DÂMASO This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8405432/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 13 Apr, 2026 Read the published version in Veterinary Research Communications → Version 1 posted 15 You are reading this latest preprint version Abstract The administration of ozone has been shown to benefit human and animal health in various areas because of its anti-inflammatory, antibacterial, antiparasitic, and antiviral effects. The effects of ozone on the health status of farm animals have not yet been described. To evaluate the clinical, haematological, and biochemical responses of sheep following rectal administration of an oxygen–ozone mixture, 10 sheep were used and divided into two experimental groups: the oxygen group (GO2), which was subjected to rectal insufflation with oxygen, and the ozone group (GO2/O3), which was subjected to rectal insufflation with an oxygen-ozone mixture. The treatments were carried out three times a week for four weeks. The animals were clinically assessed before and after all applications, and blood samples were taken at five time points. The data were analysed via analysis of variance and Tukey's test (p<0.05). There were significant differences between the groups in the number of total leucocytes and the serum concentrations of GGT, total protein, albumin, and bilirubin. The administration of the oxygen-ozone mixture had no significant effect on heart rate, red blood cell count, globular volume, haemoglobin concentration, or serum AST activity. The results showed that treatment with the oxygen-ozone mixture did not lead to significant clinical and/or haematological changes or deleterious liver changes in sheep. Therefore, ozone therapy should continue to be studied as an alternative treatment option in sheep. Haematology Liver enzymes Ozone therapy Transrectal Sheep Introduction The growing challenge in veterinary medicine of using effective and innocuous therapies that are inexpensive to perform and easy to reproduce and that reduce the use of antimicrobial agents has led to the study of ozone therapy, which is already widely used in human medicine (Pivotto et al. 2020). Ozone therapy has already demonstrated antibacterial, fungicidal, viricidal, analgesic, and anti-inflammatory effects in animal species (Sciorsci et al. 2020), promoting increased oxygen availability in the body, immune modulation, neovascularization, and tissue regeneration (Silva et al. 2018) and associated with an increase in antioxidant enzymes in the liver and kidney, intrahepatic lymphocyte activation, reduction in platelet aggregation and immunostimulant action (Kozat and Okman 2019). Ozone can be delivered pure or diluted in oils, water, or blood (Bocci 2006), and administration via intravenous, intramuscular, subcutaneous, rectal insufflation, vaginal, and intra-articular routes (Đuričić et al. 2015), with little risk of toxicity (Bocci 2006). A growing number of studies report ozone therapy for various pathologies, such as topical treatment of skin lesions, laminitis, facial nerve paralysis, lumbar pain with and without sciatica, fungal, bacterial and viral infections, with shown favourable results (Coelho et al. 2015; Ouf et al. 2016; Jaramillo et al. 2020), but evidence is still scarce on its effects in farm animals, which are commonly subjected to frequent use of antibiotics and antiparasitic drugs (Sumida and Hayashi 2022, Đuričić et al. 2015). Overall, this study aimed to investigate the effects of rectal oxygen–ozone administration on clinical, haematological, and biochemical parameters in sheep. Materials and methods This research project was approved by the Ethics Committee for the Use of Animals (CEUA) at the University of Vila Velha, in Espírito Santo, Brazil (UVV-ES), under registration number 308/2014. Ten sheep were used, including eight females and two males. The animals were clinically healthy during previous physical examinations and were subjected to the same type of diet and health management, prior to an adaptation to the experimental protocol for 30 days. The sheep were randomly divided into two experimental groups: the oxygen group (GO 2 ), composed of four females and one male (average age of 2.7 ± 1.3 years; average body weight of 41.60 ± 17.33 kg), which was subjected to rectal insufflation with oxygen gas; and the ozone group (GO 2 /O 3 ), composed of four females and one male (average age of 2.8 ± 1.3 years; average body weight of 39.80 ± 8.44 kg), which was subjected to rectal insufflation with the oxygen-ozone mixture. The calculated volume of gas to be insufflated was based on the method of Knoch et al . (1987). After the rectal ampoule was emptied, gas was applied by inserting a probe into the rectum, with oxygen alone in GO 2 and an oxygen-ozone mixture in GO 2 /O 3 . The experimental protocol (Coelho et al. 2015) was carried out over four weeks. Week 1: three applications of 20 µg/mL (300 mL) gas were made to each animal at 48-hour intervals. Week 2: the protocol was repeated (3 applications, q.48 h), changing only the concentration (25 µg/mL, 300 mL). Week 3: the concentration was maintained (25 µg/mL), and the volume was increased to 400 mL, again after 3 applications, every 48 h. Week 4: the concentration (30 µg/mL) and volume (500 mL) were increased, with 3 applications every 48 hours. The oxygen-ozone mixture was produced via an ozone generator (Ozone&Life®, São Paulo, Brazil - model 0&L 3.0 RM), which allows for oxygen flow rates ranging from 0 to 1.0 L/min (99.5% purity). The animals were assessed daily, and heart and respiratory rates, body temperature, mucous membrane color, and ruminal motility were recorded. Blood samples were taken from the jugular vein on day zero (prior to rectal administration) and once a week, 72 hours after the last weekly application, for a total of five samples per animal (T0, T1, T2, T3, and T4) to determine the liver profile (serum aspartate aminotransferase (AST), gamma glutamyl transferase (GGT), total protein, albumin, bilirubin), and blood count. The data were analysed via the GraphPad InStat 3.0 computerized statistical program, subjected to the Kolmogorov‒Smirnov normality test and considered parametric when p >0.10. The variables were subsequently subjected to repeated measures analysis of variance (ANOVA) followed by means comparison (Tukey test) with a significance level of 5%. Results The clinical assessment of the sheep in the study revealed no alterations, and they were within the normal range for the species (Seixas et al. 2021), with no alterations in rumen motility present on abdominal auscultation. Table 1 shows the mean values and standard deviations of the values recorded in the oxygen group (GO 2 ) and the ozone group (GO 2 /O 3 ) for heart rate (HR), body temperature (BT), number of red blood cells (RBC), hemoglobin concentration (Hb), globular volume (GV) and number of total leucocytes (Leuco), as well as the p values for the groups’ means comparison. Among these, changes were observed only in the total leucocyte count, with a marked increase in T1, T2, T3, and T4 to the reference standards for the species. Table 1 - Mean values and standard deviations for heart rate (HR), body temperature (HT), red blood cell count (RBC), haemoglobin concentration (Hb), globular volume (VG), and total leukocyte count (TLC) in sheep subjected to oxygen (Group O 2 ) and ozone (Group O 2 /O 3 ) insufflation at time points T0, T1, T2, T3 and T4 Group T0 T1 T2 T3 T4 p HR (bpm) O 2 73±8,2 a 75±11,5 a 74±14,3 a 66±10,8 a 82±15,1 a 0,3404 O 2 /O 3 100±15,7 a 83±22,9 a 87±19,1 a 73±15,9 a 69±16,3 a 0,094 BT ( o C) O 2 38,9±0,2 b 38,5±0,2 ab 38,0±0,5 a 38,5±0,3 ab 39,1±0,4 b 0,011 O 2 /O 3 39,2±0,5 a 38,8±0,5 b 38,5±0,5 c 38,5±0,7 c 39,3±0,4 a 0,036 RBC (x10 6 /µL) O 2 9,90±1,8 a 10,39±1,7 a 11,29± 1,4 a 10,55±2,0 a 10,67±2,0 a 0,807 O 2 /O 3 10,76±3,2 a 11,46±1,9 a 12,28±1,4 a 11,54±2,3 a 11,06±1,2 a 0,831 Hb (g/dL) O 2 10,34±1,5 a 10,94±1,1 a 12,30±1,0 a 10,82±1,5 a 11,12±1,4 a 0,250 O 2 /O 3 10,70±1,0 a 11,96±1,4 a 12,90±1,0 a 11,88±1,8 a 11,38±0,7 a 0,112 GV (%) O 2 31,6 ± 5,1 a 30,7 ± 3,2 a 37,6 ± 3,9 a 30,7 ± 4,1 a 31,4±4,1 a 0,070 O 2 /O 3 32,6 ±2,8 a 33,6 ±4,0 a 35,9±3,0 a 33,5±5,3 a 32,3±1,9 a 0,563 Leuco (/µL) O 2 9710±3821 a 22340±6478 b 23860±4358 b 22300±4363 b 18460±3384 b 0,001 O 2 /O 3 11190±2347 a 22280±2456 b 23380±4154 b 25120±3118 b 19460±3275 b <0,0001 NOTE: Lowercase letters on the same line denote a significant difference according to the Tukey test ( p <0.05). T0 (obtained before the experiment), T1 (obtained 72 hours after the first week of application), T2 (obtained 72 hours after the second week of application), T3 (obtained 72 hours after the third week of application), T4 (obtained 72 hours after the fourth week of application) Table 2 shows the mean values and standard deviations of the values in GO 2 and GO 2 /O 3 for serum AST, GGT, total protein, albumin, and bilirubin concentrations, as well as the p values determined via the Tukey test. Serum AST activity did not change in any of the two groups, and GGT did not change in the GO 2 /O 3 group. Table 2 - Mean values and standard deviations of serum total protein, albumin, and bilirubin concentrations and serum AST and GGT activities in sheep subjected to oxygen (O 2 group) and ozone (O 2 /O 3 group) insufflation at time points T0, T1, T2, T3 and T4 Group T0 T1 T2 T3 T4 p Total protein (g/dL) O 2 5,7±0,6 a 7,4±0,5 b 7,5±0,6 b 7,0±0,5 b 7,2±0,5 b <0,001 O 2 /O 3 5,7±1,3 a 7,8±0,4 b 7,8±0,4 b 7,9±0,4 b 7,7±0,7 b <0,001 Albumine (g/dL) O 2 2,8±0,4 b 1,2±0,2 a 3,1±0,1 b 3,1±0,2 b 3,1±0,2 b <0,001 O 2 /O 3 2,7±0,3 b 1,2±0,3 a 3,0±0,1 bc 3,2±0,1 c 3,1±0,2 c <0,001 AST (UI/L) O 2 136,9±57,9 a 146,3±85,3 a 129,5±51,9 a 123,5±41,7 a 120,3±33,7 a 0,946 O 2 /O 3 94,6±35,9 a 105,6±17,0 a 113,2±10,7 a 103,8±15,1 a 101,0±12,3 a 0,678 GGT (UI/L) O 2 30,7±11,6 a 41,3±6,8 b 38,3±4,4 ab 37,8±5,7 ab 37,0±6,0 ab 0,011 O 2 /O 3 34,8±6,3 a 45,6±7,8 a 41,0±9,8 a 44,0±8,9 a 36,0±12,3 a 0,059 Total bilirrubine (mg/dL) O 2 0,26 ± 0,06 b 0,11 ± 0,06 a 0,11 ± 0,06 a 0,13 ± 0,06 a 0,19±0,02 ab 0,003 O 2 /O 3 0,28 ± 0,07 b 0,09 ± 0,07 a 0,09 ± 0,06 a 0,11 ± 0,06 a 0,13 ± 0,06 a <0,001 NOTE: Lowercase letters on the same line denote a significant difference according to the Tukey test ( p <0.05). T0 (obtained before the experiment), T1 (obtained after 72 hours of the first week of application), T2 (obtained after 72 hours of the second week of application), T3 (obtained after 72 hours of the third week of application), T4 (obtained after 72 hours of the fourth week of application). Discussion In this study, an initial dose of 20 µg/mL was established as the oxidative preconditioning protocol, since doses of less than 10 µg/mL are considered biologically inefficient because they do not reach the therapeutic window owing to neutralization by circulating serum antioxidant enzymes (Sagai and Bocci 2011; Coelho et al. 2015). In addition, the doses selected were within the range of 10 to 80 µg/mL, which are considered beneficial because they promote immunomodulation, favour the activation of the endogenous antioxidant system via induced mild and transient oxidative stress, and help prevent hepatocellular lesions caused by free radicals (Menendez-Cepero et al. 2018). The clinical assessment of the sheep used showed no significant changes, with the values obtained being within the normal range for the species (Seixas et al. 2021). Only the average body temperature was close to the upper limit in both groups and at all assessment times, which may be related to the tropical climate of the region (daSilva and Minomo 1995). In addition, sudden restraint can trigger an alert reaction in animals, which is responsible for increases in heart and respiratory rates and body temperature (Grandin and Shivley 2015). However, during the same periods, there was no change in the HR. In general, the results observed in the physical parameters were similar to the outcome in horses subjected to the therapeutic use of ozone via the transrectal route at concentrations of 10, 15, and 20 µg/mL (Coelho et al. 2015). Similarly, exposure of healthy horses to up to 1000 mL of the therapeutic ozone mixture by intravenous administration did not cause clinical changes in the same physical parameters mentioned above (Sciorsci et al. 2020). Several studies have reported ozone therapy in ruminants to treat different disorders, including metritis and endometritis (Maldonado et al. 2017), urovagina (Zobel et al. 2012), and retained placenta (Zobel et al. 2013), but no clinical parameters have been described in these studies to allow comparisons with the current ones. The use of major autohaemotherapy in calves to assess the influence on immunological values revealed that treatment with ozone gas promoted leucocytosis (Terasaki et al. 2001). Similarly, in the present study, leucocytosis was observed in both groups from the first rectal insufflation onwards. A possible and important cause of the increase in the number of leucocytes could be the stress caused by handling and restraining the animals for sample collection, although these animals were previously subjected to manipulation and discomfort or even pain following rectal administration, as stress promotes the endogenous release of glucocorticoids, increasing blood circulation so that leucocytes can move to the peripheral blood (Tennant 2008). Another possibility is that immune cells can take advantage of the oxygen availability generated by O 3 for metabolism (Bocci 2006). On the other hand, in horses, rectal insufflation did not promote changes in the number of leucocytes (Coelho et al. 2015), which was explained by the authors with healthy horses that underwent no clinical challenge during the experiment. The values recorded throughout the experimental period for the biomarkers selected were within the reference range for sheep (Tennant 2008), except for albumin, whose results at T1 (average of 1.2 mg/dL for GO 2 and GO 2 /O 3 ) were lower than the normal range described (2.4–3.0 mg/dL). Hypoalbuminemia is observed in cases of chronic and severe hepatopathies and is associated with an increase in globulins and, concomitantly, an increase in total serum proteins (Sumida and Hayashi 2022). Hypoalbuminemia has been associated to failure in its synthesis because of chronic liver diseases, gastrointestinal diseases, or losses due to nephropathy (Tennant 2008). In this study, for both groups, a significant reduction in albumin (hypoalbuminemia) was associated with a significant increase in total protein and possibly hyperglobulinemia in the first week after rectal insufflation. This could have been due to diet and nutritional adaptations to feed, especially forage, which is a feedstuff that is mandatory for ruminants but difficult to maintain a constant nutritional content. From the second week of treatment, the albumin values normalized, whereas the total serum protein values remained at the same level until the last assessment point (T4) and remained within the normal range (Tennant 2008). Furthermore, in GO 2, there was a transient increase in GGT at the same time. GGT is a hepatic indicator that can be elevated in the case of cholestasis (Tennant 2008), which can be explained by reduced food intake. The latter could be associated with the foodstuff available at the time, which, as mentioned before, was not intentionally changed, or perhaps was due to discomfort in the rectum due to the treatment administration. This change in liver function was transitory, due to the subsequent recovery of albumin and GGT values and maintenance of total protein values, all within the normal range for the species (Tennant 2008). The significant reduction in total bilirubin at T1 may be correlated with hypoalbuminemia at T1 since the unconjugated portion of bilirubin (indirect bilirubin) is transported by plasma albumin to the liver (Tennant 2008). In line with the beneficial effects already described in veterinary patients, such as antibacterial (Maldonado et al. 2017), reduction of chronic (Coelho et al. 2015) and acute (Rodríguez et al. 2019) inflammation, this work seeks to contribute to the understanding of the effects, as well as possible complications in the use of medical ozone in sheep, through biomarkers, such as those described in horses (Jaramillo et al. 2020). Conclusion Treatment with the oxygen-ozone mixture at increasing doses from 20 to 30 µg/mL proved to be safe and did not cause significant changes in biomarkers that could signal deleterious lesions induced by sheep therapy. However, as this is a new therapy, further research needs to be carried out with different doses and concentrations to assess both its therapeutic efficacy and possible harmful effects on different animal species. Declarations Authors’ contributions VINICIUS R.C. SOUZA: Experimental design, application for approval of the ethical committee, data collection and data analysis. RENAN S. CARVALHO: Data collection and data analysis. CLARISSE S. COELHO: Experimental design and drafting of the manuscript. ÂNGELA DÂMASO: Major review of the work for important intellectual content. Conflict of interest: The authors have no conflicts of interest to disclose. References Bocci AV. Scientific and Medical Aspects of Ozone Therapy. State of the Art. Review article Department of Physiology, Archives Med Res (UniSi) 2006:37:425-35. Coelho CS, Abreu-Bernadi W, Ginelli AM, et al. Use of ozone therapy in chronic laminitis in a horse. J Ozone Therapy 2015:1:30-5. daSilva RG, Minomo FR. Circadian and seasonal variation of the body temperature of sheep in a tropical environment. Int J Biometeorol, 1995:39:69–73. Đuričić D, Valpotić H, Samardžija M. Prophylaxis and therapeutic potential of ozone in buiatrics: Current knowledge. Anim Reprod Sci 2015:159:1-7. Grandin T, Shivley C. How Farm Animals React and Perceive Stressful Situations Such As Handling, Restraint, and Transport. Animals 2015:5:1233-51 Jaramillo FM, Vendruscolo CP, Fülber J, et al. Effects of transrectal medicinal ozone in horses - clinical and laboratory aspects. Arq Bras Med Vet Zootec 2020:72:56-64. Kozat S, Okman E. Has Ozone Therapy a Future in Veterinary Medicine? J Animal Husbandry & Dairy Science 2019:3:25-34. Maldonado H, Narváez J, Enríquez M, Ortuño C. 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Seixas L, Peripolli V, Façanha DAE, et al. Physiological and hematological parameters of sheep reared in the tropics and subtropics. Arq Bras Med Vet Zootec 2021:73:622-30 Silva TC da, Shiosi RK, Raineri Neto R. Ozonioterapia: um tratamento clínico em ascensão na medicina veterinária - revisão de literatura. Rev Cient Med Vet 2018:31:1679-7353 Sumida JM, Hayashi AM. Ozone therapy in veterinary medicine: clinical indications and techniques. Acta Vet Bras 2022: 16:294-304 Tennant BC. Hepatic Function. In: Kaneko, J. J.; Harvey, J. W.; Bruss, M. C. Clinical Biochemistry of Domestic Animals. 6 th ed. San Diego: Academic Pres: 2008. Terasaki N, Ogata A, Ohtsuka H. Changes of immunological responses after experimentally ozonated administration in calves, J Vet Med Sci 2001:63:1327-30. Zobel R, Tkalčić S, Štoković I, et al. Efficacy of Ozone as a Novel Treatment Option for Urovagina in Dairy Cows. Reprod Domest Anim = Zuchthygiene; 2012:47:293–8. Zobel R, Tkalčić S (2013) Efficacy of ozone and other treatment modalities for retained placenta in dairy cows. Reprod Domest Anim = Zuchthygiene 2013:48:121–5. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 13 Apr, 2026 Read the published version in Veterinary Research Communications → Version 1 posted Editorial decision: Revision requested 08 Jan, 2026 Reviews received at journal 07 Jan, 2026 Reviews received at journal 04 Jan, 2026 Reviews received at journal 03 Jan, 2026 Reviews received at journal 02 Jan, 2026 Reviewers agreed at journal 29 Dec, 2025 Reviewers agreed at journal 25 Dec, 2025 Reviewers agreed at journal 25 Dec, 2025 Reviewers agreed at journal 23 Dec, 2025 Reviewers agreed at journal 23 Dec, 2025 Reviewers agreed at journal 23 Dec, 2025 Reviewers invited by journal 23 Dec, 2025 Editor assigned by journal 23 Dec, 2025 Submission checks completed at journal 23 Dec, 2025 First submitted to journal 19 Dec, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8405432","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":564794603,"identity":"163ba4a4-7b3d-4783-8e29-6a9085664f2f","order_by":0,"name":"VINICIUS R.C. SOUZA","email":"","orcid":"","institution":"Lusófona University - Lisbon University Center","correspondingAuthor":false,"prefix":"","firstName":"VINICIUS","middleName":"R.C.","lastName":"SOUZA","suffix":""},{"id":564794605,"identity":"8e6961a4-7354-49af-b8e2-8d9e2cd40b6c","order_by":1,"name":"RENAN S. CARVALHO","email":"","orcid":"","institution":"University of Vila Velha","correspondingAuthor":false,"prefix":"","firstName":"RENAN","middleName":"S.","lastName":"CARVALHO","suffix":""},{"id":564794606,"identity":"155b5aa5-92cd-479a-b6a1-c5616e24af5d","order_by":2,"name":"CLARISSE S. COELHO","email":"","orcid":"","institution":"CECAV - Lusófona University","correspondingAuthor":false,"prefix":"","firstName":"CLARISSE","middleName":"S.","lastName":"COELHO","suffix":""},{"id":564794607,"identity":"25cb39d4-303f-4888-8776-c6a6fe7f2e5d","order_by":3,"name":"ÂNGELA DÂMASO","email":"data:image/png;base64,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","orcid":"","institution":"Lusófona University - Lisbon University Center","correspondingAuthor":true,"prefix":"","firstName":"ÂNGELA","middleName":"","lastName":"DÂMASO","suffix":""}],"badges":[],"createdAt":"2025-12-19 13:53:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8405432/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8405432/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11259-026-11205-4","type":"published","date":"2026-04-13T15:59:33+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":98998613,"identity":"6cbffac3-c1a8-4561-aba3-c20a7e5a0ac3","added_by":"auto","created_at":"2025-12-25 13:36:09","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":37866,"visible":true,"origin":"","legend":"","description":"","filename":"BriefReportClinicalandbloodparametersozonoterapyinsheepVetResComunicationsDec2025.docx","url":"https://assets-eu.researchsquare.com/files/rs-8405432/v1/e38e4cb578e0bd2a4f179efa.docx"},{"id":98998612,"identity":"450f56ed-9243-43fb-a5a7-0299a17a6bb5","added_by":"auto","created_at":"2025-12-25 13:36:09","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":5853,"visible":true,"origin":"","legend":"","description":"","filename":"4ec4df47ec714188bb00a9cb4d30cded.json","url":"https://assets-eu.researchsquare.com/files/rs-8405432/v1/08ca625ad5cd3b8ff2d20764.json"},{"id":99311983,"identity":"621ea854-c1a0-44b7-8012-bbddede832d0","added_by":"auto","created_at":"2025-12-31 16:17:31","extension":"xml","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":71944,"visible":true,"origin":"","legend":"","description":"","filename":"4ec4df47ec714188bb00a9cb4d30cded1enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-8405432/v1/c17073ced9672282f3b9d282.xml"},{"id":98998615,"identity":"8c6ac26d-5eee-4f15-a735-f605d54a545a","added_by":"auto","created_at":"2025-12-25 13:36:09","extension":"xml","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":68326,"visible":true,"origin":"","legend":"","description":"","filename":"4ec4df47ec714188bb00a9cb4d30cded1structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-8405432/v1/bc2fd173485fc5f3310d4872.xml"},{"id":98998616,"identity":"bc3af14a-f70b-4ac0-9eaa-9adec3f59031","added_by":"auto","created_at":"2025-12-25 13:36:09","extension":"html","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":76507,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8405432/v1/5397850d1a9560ff890d4599.html"},{"id":107351077,"identity":"56ea1b28-bdf3-4ca2-ad8a-bc5f36cc8030","added_by":"auto","created_at":"2026-04-20 16:09:05","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":326295,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8405432/v1/9171fe55-43c3-4340-b665-3bfe52b63418.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Clinical, haematological, and biochemical responses of rectal oxygen-ozone administration in sheep","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe growing challenge in veterinary medicine of using effective and innocuous therapies that are inexpensive to perform and easy to reproduce and that reduce the use of antimicrobial agents has led to the study of ozone therapy, which is already widely used in human medicine (Pivotto et al. 2020). Ozone therapy has already demonstrated antibacterial, fungicidal, viricidal, analgesic, and anti-inflammatory effects in animal species (Sciorsci et al. 2020), promoting increased oxygen availability in the body, immune modulation, neovascularization, and tissue regeneration (Silva et al. 2018) and associated with an increase in antioxidant enzymes in the liver and kidney, intrahepatic lymphocyte activation, reduction in platelet aggregation and immunostimulant action (Kozat and Okman 2019).\u003c/p\u003e\n\u003cp\u003eOzone can be delivered pure or diluted in oils, water, or blood (Bocci 2006), and administration via intravenous, intramuscular, subcutaneous, rectal insufflation, vaginal, and intra-articular routes (Đuričić et al. 2015), with little risk of toxicity (Bocci 2006). A growing number of studies report ozone therapy for various pathologies, such as topical treatment of skin lesions, laminitis, facial nerve paralysis, lumbar pain with and without sciatica, fungal, bacterial and viral infections, with shown favourable results (Coelho et al. 2015; Ouf et al. 2016; Jaramillo et al. 2020), but evidence is still scarce on its effects in farm animals, which are commonly subjected to frequent use of antibiotics and antiparasitic drugs (Sumida and Hayashi 2022,\u0026nbsp;Đuričić et al. 2015).\u003c/p\u003e\n\u003cp\u003eOverall, this study aimed to investigate the effects of rectal oxygen\u0026ndash;ozone administration on clinical, haematological, and biochemical parameters in sheep.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003eThis research project was approved by the Ethics Committee for the Use of Animals (CEUA) at the University of Vila Velha, in Esp\u0026iacute;rito Santo, Brazil (UVV-ES), under registration number 308/2014.\u003c/p\u003e\n\u003cp\u003eTen sheep were used, including eight females and two males. The animals were clinically healthy during previous physical examinations and were subjected to the same type of diet and health management, prior to an adaptation to the experimental protocol for 30 days.\u003c/p\u003e\n\u003cp\u003eThe sheep were randomly divided into two experimental groups: the oxygen group (GO\u003csub\u003e2\u003c/sub\u003e), composed of four females and one male (average age of 2.7 \u0026plusmn; 1.3 years; average body weight of 41.60 \u0026plusmn; 17.33 kg), which was subjected to rectal insufflation with oxygen gas; and the ozone group (GO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e), composed of four females and one male (average age of 2.8 \u0026plusmn; 1.3 years; average body weight of 39.80 \u0026plusmn; 8.44 kg), which was subjected to rectal insufflation with the oxygen-ozone mixture.\u003c/p\u003e\n\u003cp\u003eThe calculated volume of gas to be insufflated was based on the method of Knoch et al\u003cem\u003e.\u003c/em\u003e (1987). After the rectal ampoule was emptied, gas was applied by inserting a probe into the rectum, with oxygen alone in GO\u003csub\u003e2\u003c/sub\u003e and an oxygen-ozone mixture in GO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e.\u003c/p\u003e\n\u003cp\u003eThe experimental protocol (Coelho et al. 2015) was carried out over four weeks. Week 1: three applications of 20 \u0026micro;g/mL (300 mL) gas were made to each animal at 48-hour intervals. Week 2: the protocol was repeated (3 applications, q.48 h), changing only the concentration (25 \u0026micro;g/mL, 300 mL). Week 3: the concentration was maintained (25 \u0026micro;g/mL), and the volume was increased to 400 mL, again after 3 applications, every 48 h. Week 4: the concentration (30 \u0026micro;g/mL) and volume (500 mL) were increased, with 3 applications every 48 hours.\u003c/p\u003e\n\u003cp\u003eThe oxygen-ozone mixture was produced via an ozone generator (Ozone\u0026amp;Life\u0026reg;, S\u0026atilde;o Paulo, Brazil - model 0\u0026amp;L 3.0 RM), which allows for oxygen flow rates ranging from 0 to 1.0 L/min (99.5% purity).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe animals were assessed daily, and heart and respiratory rates, body temperature, mucous membrane color, and ruminal motility were recorded. Blood samples were taken from the jugular vein on day zero (prior to rectal administration) and once a week, 72 hours after the last weekly application, for a total of five samples per animal (T0, T1, T2, T3, and T4) to determine the liver profile (serum aspartate aminotransferase (AST), gamma glutamyl transferase (GGT), total protein, albumin, bilirubin), and blood count.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe data were analysed via the GraphPad InStat 3.0 computerized statistical program, subjected to the Kolmogorov‒Smirnov normality test and considered parametric when \u003cem\u003ep\u003c/em\u003e\u0026gt;0.10. The variables were subsequently subjected to repeated measures analysis of variance (ANOVA) followed by means comparison (Tukey test) with a significance level of 5%.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe clinical assessment of the sheep in the study revealed no alterations, and they were within the normal range for the species (Seixas et al. 2021), with no alterations in rumen motility present on abdominal auscultation. Table 1 shows the mean values and standard deviations of the values recorded in the oxygen group (GO\u003csub\u003e2\u003c/sub\u003e) and the ozone group (GO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e) for heart rate (HR), body temperature (BT), number of red blood cells (RBC), hemoglobin concentration (Hb), globular volume (GV) and number of total leucocytes (Leuco), as well as the \u003cem\u003ep\u003c/em\u003e values for the groups\u0026rsquo; means comparison. Among these, changes were observed only in the total leucocyte count, with a marked increase in T1, T2, T3, and T4 to the reference standards for the species.\u003c/p\u003e\n\u003cp\u003eTable 1 - Mean values and standard deviations for heart rate (HR), body temperature (HT), red blood cell count (RBC), haemoglobin concentration (Hb), globular volume (VG), and total leukocyte count (TLC) in sheep subjected to oxygen (Group O\u003csub\u003e2\u003c/sub\u003e) and ozone (Group O\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e) insufflation at time points T0, T1, T2, T3 and T4\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"576\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eGroup\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003eT0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003eT1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003eT2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 83px;\"\u003e\n \u003cp\u003eT3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003eT4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eHR (bpm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e73\u0026plusmn;8,2\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e75\u0026plusmn;11,5\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e74\u0026plusmn;14,3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e66\u0026plusmn;10,8\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e82\u0026plusmn;15,1\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,3404\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e100\u0026plusmn;15,7\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e83\u0026plusmn;22,9\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e87\u0026plusmn;19,1\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e73\u0026plusmn;15,9\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e69\u0026plusmn;16,3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,094\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eBT (\u003csup\u003eo\u003c/sup\u003eC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e38,9\u0026plusmn;0,2\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e38,5\u0026plusmn;0,2\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e38,0\u0026plusmn;0,5\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e38,5\u0026plusmn;0,3\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e39,1\u0026plusmn;0,4\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,011\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e39,2\u0026plusmn;0,5\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e38,8\u0026plusmn;0,5\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e38,5\u0026plusmn;0,5\u003csup\u003e\u0026nbsp;c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e38,5\u0026plusmn;0,7\u003csup\u003e\u0026nbsp;c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e39,3\u0026plusmn;0,4\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,036\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eRBC (x10\u003csup\u003e6\u003c/sup\u003e/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e9,90\u0026plusmn;1,8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,39\u0026plusmn;1,7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,29\u0026plusmn;\u0026nbsp;1,4\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,55\u0026plusmn;2,0\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,67\u0026plusmn;2,0\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,807\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,76\u0026plusmn;3,2\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,46\u0026plusmn;1,9\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e12,28\u0026plusmn;1,4\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,54\u0026plusmn;2,3\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,06\u0026plusmn;1,2\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,831\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eHb (g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,34\u0026plusmn;1,5\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,94\u0026plusmn;1,1\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e12,30\u0026plusmn;1,0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,82\u0026plusmn;1,5\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,12\u0026plusmn;1,4\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,250\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e10,70\u0026plusmn;1,0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,96\u0026plusmn;1,4\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e12,90\u0026plusmn;1,0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,88\u0026plusmn;1,8\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11,38\u0026plusmn;0,7\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,112\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eGV (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e31,6\u0026nbsp;\u0026plusmn;\u0026nbsp;5,1\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e30,7\u0026nbsp;\u0026plusmn;\u0026nbsp;3,2\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e37,6\u0026nbsp;\u0026plusmn;\u0026nbsp;3,9\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e30,7\u0026nbsp;\u0026plusmn;\u0026nbsp;4,1\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e31,4\u0026plusmn;4,1\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,070\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e32,6\u0026nbsp;\u0026plusmn;2,8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e33,6\u0026nbsp;\u0026plusmn;4,0\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e35,9\u0026plusmn;3,0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e33,5\u0026plusmn;5,3\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e32,3\u0026plusmn;1,9\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,563\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eLeuco (/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e9710\u0026plusmn;3821\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e22340\u0026plusmn;6478\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e23860\u0026plusmn;4358\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e22300\u0026plusmn;4363\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e18460\u0026plusmn;3384\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11190\u0026plusmn;2347\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e22280\u0026plusmn;2456\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e23380\u0026plusmn;4154\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e25120\u0026plusmn;3118\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e19460\u0026plusmn;3275\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0,0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 67px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 80px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 59px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eNOTE: Lowercase letters on the same line denote a significant difference according to the Tukey test (\u003cem\u003ep\u003c/em\u003e\u0026lt;0.05). T0 (obtained before the experiment), T1 (obtained 72 hours after the first week of application), T2 (obtained 72 hours after the second week of application), T3 (obtained 72 hours after the third week of application), T4 (obtained 72 hours after the fourth week of application)\u003c/p\u003e\n\u003cp\u003eTable 2 shows the mean values and standard deviations of the values in GO\u003csub\u003e2\u003c/sub\u003e and GO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e for serum AST, GGT, total protein, albumin, and bilirubin concentrations, as well as the \u003cem\u003ep\u003c/em\u003e values determined via the Tukey test. Serum AST activity did not change in any of the two groups, and GGT did not change in the GO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u0026nbsp;\u003c/sub\u003egroup.\u003c/p\u003e\n\u003cp\u003eTable 2 - Mean values and standard deviations of serum total protein, albumin, and bilirubin concentrations and serum AST and GGT activities in sheep subjected to oxygen (O\u003csub\u003e2\u003c/sub\u003e group) and ozone (O\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e group) insufflation at time points T0, T1, T2, T3 and T4\u003c/p\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"559\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eGroup\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eT0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eT1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eT2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eT3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eT4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eTotal protein (g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e5,7\u0026plusmn;0,6\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,4\u0026plusmn;0,5\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,5\u0026plusmn;0,6\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,0\u0026plusmn;0,5\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,2\u0026plusmn;0,5\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0,001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e5,7\u0026plusmn;1,3\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,8\u0026plusmn;0,4\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,8\u0026plusmn;0,4\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,9\u0026plusmn;0,4\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e7,7\u0026plusmn;0,7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0,001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eAlbumine (g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e2,8\u0026plusmn;0,4\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e1,2\u0026plusmn;0,2\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3,1\u0026plusmn;0,1\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3,1\u0026plusmn;0,2\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3,1\u0026plusmn;0,2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0,001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e2,7\u0026plusmn;0,3\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e1,2\u0026plusmn;0,3\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3,0\u0026plusmn;0,1\u003csup\u003e\u0026nbsp;bc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3,2\u0026plusmn;0,1\u003csup\u003e\u0026nbsp;c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3,1\u0026plusmn;0,2\u003csup\u003e\u0026nbsp;c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0,001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eAST (UI/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e136,9\u0026plusmn;57,9\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e146,3\u0026plusmn;85,3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e129,5\u0026plusmn;51,9\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e123,5\u0026plusmn;41,7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e120,3\u0026plusmn;33,7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,946\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e94,6\u0026plusmn;35,9\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e105,6\u0026plusmn;17,0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e113,2\u0026plusmn;10,7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e103,8\u0026plusmn;15,1\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e101,0\u0026plusmn;12,3\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,678\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eGGT (UI/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e30,7\u0026plusmn;11,6\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e41,3\u0026plusmn;6,8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e38,3\u0026plusmn;4,4\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e37,8\u0026plusmn;5,7\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e37,0\u0026plusmn;6,0\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,011\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e34,8\u0026plusmn;6,3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e45,6\u0026plusmn;7,8\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e41,0\u0026plusmn;9,8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e44,0\u0026plusmn;8,9\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e36,0\u0026plusmn;12,3\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e0,059\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003eTotal bilirrubine (mg/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,26\u0026nbsp;\u0026plusmn;\u0026nbsp;0,06\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,11\u0026nbsp;\u0026plusmn;\u0026nbsp;0,06\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,11\u0026nbsp;\u0026plusmn;\u0026nbsp;0,06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,13\u0026nbsp;\u0026plusmn;\u0026nbsp;0,06\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,19\u0026plusmn;0,02\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,003\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,28\u0026nbsp;\u0026plusmn;\u0026nbsp;0,07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,09\u0026nbsp;\u0026plusmn;\u0026nbsp;0,07\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,09\u0026nbsp;\u0026plusmn;\u0026nbsp;0,06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,11\u0026nbsp;\u0026plusmn;\u0026nbsp;0,06\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0,13\u0026nbsp;\u0026plusmn;\u0026nbsp;0,06\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0,001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 70px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 77px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 77px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 77px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 77px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 77px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eNOTE: Lowercase letters on the same line denote a significant difference according to the Tukey test (\u003cem\u003ep\u003c/em\u003e\u0026lt;0.05). T0 (obtained before the experiment), T1 (obtained after 72 hours of the first week of application), T2 (obtained after 72 hours of the second week of application), T3 (obtained after 72 hours of the third week of application), T4 (obtained after 72 hours of the fourth week of application).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this study, an initial dose of 20 \u0026micro;g/mL was established as the oxidative preconditioning protocol, since doses of less than 10 \u0026micro;g/mL are considered biologically inefficient because they do not reach the therapeutic window owing to neutralization by circulating serum antioxidant enzymes (Sagai and Bocci 2011; Coelho et al. 2015). In addition, the doses selected were within the range of 10 to 80 \u0026micro;g/mL, which are considered beneficial because they promote immunomodulation, favour the activation of the endogenous antioxidant system via induced mild and transient oxidative stress, and help prevent hepatocellular lesions caused by free radicals (Menendez-Cepero et al. 2018).\u003c/p\u003e\n\u003cp\u003eThe clinical assessment of the sheep used showed no significant changes, with the values obtained being within the normal range for the species (Seixas et al. 2021). Only the average body temperature was close to the upper limit in both groups and at all assessment times, which may be related to the tropical climate of the region (daSilva and Minomo 1995). In addition, sudden restraint can trigger an alert reaction in animals, which is responsible for increases in heart and respiratory rates and body temperature (Grandin and Shivley 2015). However, during the same periods, there was no change in the HR.\u003c/p\u003e\n\u003cp\u003eIn general, the results observed in the physical parameters were similar to the outcome in horses subjected to the therapeutic use of ozone via the transrectal route at concentrations of 10, 15, and 20\u0026nbsp;\u0026micro;g/mL (Coelho et al. 2015). Similarly, exposure of healthy horses to up to 1000 mL of the therapeutic ozone mixture by intravenous administration did not cause clinical changes in the same physical parameters mentioned above (Sciorsci et al. 2020). Several studies have reported ozone therapy in ruminants to treat different disorders, including metritis and endometritis (Maldonado et al. 2017), urovagina (Zobel et al. 2012), and retained placenta (Zobel et al. 2013), but no clinical parameters have been described in these studies to allow comparisons with the current ones.\u003c/p\u003e\n\u003cp\u003eThe use of major autohaemotherapy in calves to assess the influence on immunological values revealed that treatment with ozone gas promoted leucocytosis (Terasaki et al. 2001). Similarly, in the present study, leucocytosis was observed in both groups from the first rectal insufflation onwards. A possible and important cause of the increase in the number of leucocytes could be the stress caused by handling and restraining the animals for sample collection, although these animals were previously subjected to manipulation and discomfort or even pain following rectal administration, as stress promotes the endogenous release of glucocorticoids, increasing blood circulation so that leucocytes can move to the peripheral blood (Tennant 2008). Another possibility is that immune cells can take advantage of the oxygen availability generated by O\u003csub\u003e3\u003c/sub\u003e for metabolism (Bocci 2006). On the other hand, in horses, rectal insufflation did not promote changes in the number of leucocytes (Coelho et al. 2015), which was explained by the authors with healthy horses that underwent no clinical challenge during the experiment.\u003c/p\u003e\n\u003cp\u003eThe values recorded throughout the experimental period for the biomarkers selected were within the reference range for sheep (Tennant 2008), except for albumin, whose results at T1 (average of 1.2 mg/dL for GO\u003csub\u003e2\u003c/sub\u003e and GO\u003csub\u003e2\u003c/sub\u003e/O\u003csub\u003e3\u003c/sub\u003e) were lower than the normal range described (2.4\u0026ndash;3.0 mg/dL). Hypoalbuminemia is observed in cases of chronic and severe hepatopathies and is associated with an increase in globulins and, concomitantly, an increase in total serum proteins (Sumida and Hayashi 2022). Hypoalbuminemia has been associated to failure in its synthesis because of chronic liver diseases, gastrointestinal diseases, or losses due to nephropathy (Tennant 2008). In this study, for both groups, a significant reduction in albumin (hypoalbuminemia) was associated with a significant increase in total protein and possibly hyperglobulinemia in the first week after rectal insufflation. This could have been due to diet and nutritional adaptations to feed, especially forage, which is a feedstuff that is mandatory for ruminants but difficult to maintain a constant nutritional content. From the second week of treatment, the albumin values normalized, whereas the total serum protein values remained at the same level until the last assessment point (T4) and remained within the normal range (Tennant 2008). Furthermore, in GO\u003csub\u003e2,\u003c/sub\u003e there was a transient increase in GGT at the same time. GGT is a hepatic indicator that can be elevated in the case of cholestasis (Tennant 2008), which can be explained by reduced food intake. The latter could be associated with the foodstuff available at the time, which, as mentioned before, was not intentionally changed, or perhaps was due to discomfort in the rectum due to the treatment administration. This change in liver function was transitory, due to the subsequent recovery of albumin and GGT values and maintenance of total protein values, all within the normal range for the species (Tennant 2008). The significant reduction in total bilirubin at T1 may be correlated with hypoalbuminemia at T1 since the unconjugated portion of bilirubin (indirect bilirubin) is transported by plasma albumin to the liver (Tennant 2008).\u003c/p\u003e\n\u003cp\u003eIn line with the beneficial effects already described in veterinary patients, such as antibacterial (Maldonado et al. 2017), reduction of chronic (Coelho et al. 2015) and acute (Rodr\u0026iacute;guez et al. 2019) inflammation, this work seeks to contribute to the understanding of the effects, as well as possible complications in the use of medical ozone in sheep, through biomarkers, such as those described in horses (Jaramillo et al. 2020).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eTreatment with the oxygen-ozone mixture at increasing doses from 20 to 30 µg/mL proved to be safe and did not cause significant changes in biomarkers that could signal deleterious lesions induced by sheep therapy. However, as this is a new therapy, further research needs to be carried out with different doses and concentrations to assess both its therapeutic efficacy and possible harmful effects on different animal species.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eVINICIUS R.C. SOUZA:\u0026nbsp;\u003c/strong\u003eExperimental design, application for approval of the ethical committee, data collection and data analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRENAN S. CARVALHO:\u0026nbsp;\u003c/strong\u003eData collection and data analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCLARISSE S. COELHO:\u0026nbsp;\u003c/strong\u003eExperimental design and drafting of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026Acirc;NGELA D\u0026Acirc;MASO:\u0026nbsp;\u003c/strong\u003eMajor review of the work for important intellectual content.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest:\u003c/strong\u003e The authors have no conflicts of interest to disclose.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eBocci AV. Scientific and Medical Aspects of Ozone Therapy. State of the Art. Review article Department of Physiology, Archives Med Res (UniSi) 2006:37:425-35.\u003c/li\u003e\n \u003cli\u003eCoelho CS, Abreu-Bernadi W, Ginelli AM, et al. Use of ozone therapy in chronic laminitis in a horse. J Ozone Therapy 2015:1:30-5.\u003c/li\u003e\n \u003cli\u003edaSilva RG, Minomo FR. Circadian and seasonal variation of the body temperature of sheep in a tropical environment.\u0026nbsp;Int J Biometeorol,\u0026nbsp;1995:39:69\u0026ndash;73.\u003c/li\u003e\n \u003cli\u003eĐuričić D, Valpotić H, Samardžija M. Prophylaxis and therapeutic potential of ozone in buiatrics: Current knowledge. Anim Reprod Sci 2015:159:1-7.\u003c/li\u003e\n \u003cli\u003eGrandin T, Shivley C. How Farm Animals React and Perceive Stressful Situations Such As Handling, Restraint, and Transport. Animals 2015:5:1233-51\u003c/li\u003e\n \u003cli\u003eJaramillo FM, Vendruscolo CP, F\u0026uuml;lber J, et al. Effects of transrectal medicinal ozone in horses - clinical and laboratory aspects.\u0026nbsp;Arq Bras Med Vet Zootec 2020:72:56-64.\u003c/li\u003e\n \u003cli\u003eKozat S, Okman E. Has Ozone Therapy a Future in Veterinary Medicine? J Animal Husbandry \u0026amp; Dairy Science 2019:3:25-34.\u003c/li\u003e\n \u003cli\u003eMaldonado H, Narv\u0026aacute;ez J, Enr\u0026iacute;quez M, Ortu\u0026ntilde;o C. Uso de la ozonoterapia para el control de la endometritis subcl\u0026iacute;nica postparto en vacas lecheras. Maskana\u003cem\u003e\u0026nbsp;\u003c/em\u003e2017:8:105\u0026ndash;8.\u003c/li\u003e\n \u003cli\u003eMenendez-Cepero S, Leon OS, Martinez G, Alvarez H. Ozone Therapy in diabetes and its complications.\u0026nbsp;J Ozone Therapy 2018:2.\u003c/li\u003e\n \u003cli\u003eOuf SA, Moussa TA, Abd-Elmegeed AM, Eltahlawy SR. Anti-fungal potential of ozone against some dermatophytes. Braz J Microbiol 2016:47:697-702.\u003c/li\u003e\n \u003cli\u003ePivotto AP, Banhuk WF, Staffen IV, et al. Clinical uses and molecular aspects of ozone therapy: a review. OnLine J Biol Sci 2020:20:37-49.\u003c/li\u003e\n \u003cli\u003eRodr\u0026iacute;guez ZZ, Mart\u0026iacute;nez AG, Napoles LO, et al. Evaluation of the dermal and ophthalmic irritability of the ozonized sunflower oil formulation in rabbits. Revista CENIC 2019:50:242\u0026ndash;53.\u003c/li\u003e\n \u003cli\u003eSagai M, Bocci V. Mechanisms of action involved in ozone therapy: is healing induced via a mild oxidative stress? Med Gas Res 2011:1:29.\u003c/li\u003e\n \u003cli\u003eSciorsci RL, Lillo E, Occhiogrosso L, Rizzo A. Ozone therapy in veterinary medicine: A review. Res Vet Sci 2020:130:240\u0026ndash;6.\u003c/li\u003e\n \u003cli\u003eSeixas L, Peripolli V, Fa\u0026ccedil;anha DAE, et al. Physiological and hematological parameters of sheep reared in the tropics and subtropics. Arq Bras Med Vet Zootec 2021:73:622-30\u003c/li\u003e\n \u003cli\u003eSilva TC da, Shiosi RK, Raineri Neto R. Ozonioterapia: um tratamento cl\u0026iacute;nico em ascens\u0026atilde;o na medicina veterin\u0026aacute;ria - revis\u0026atilde;o de literatura. Rev Cient Med Vet 2018:31:1679-7353\u003c/li\u003e\n \u003cli\u003eSumida JM, Hayashi AM.\u0026nbsp;Ozone therapy in veterinary medicine: clinical indications and techniques. Acta Vet Bras 2022: 16:294-304\u003c/li\u003e\n \u003cli\u003eTennant BC. Hepatic Function. In: Kaneko, J. J.; Harvey, J. W.; Bruss, M. C. Clinical Biochemistry of Domestic Animals. 6\u003csup\u003eth\u003c/sup\u003e ed. San Diego: Academic Pres: 2008.\u003c/li\u003e\n \u003cli\u003eTerasaki N, Ogata A, Ohtsuka H. Changes of immunological responses after experimentally ozonated administration in calves, J Vet Med Sci 2001:63:1327-30.\u003c/li\u003e\n \u003cli\u003eZobel R, Tkalčić S, \u0026Scaron;toković I, et al. Efficacy of Ozone as a Novel Treatment Option for Urovagina in Dairy Cows. Reprod Domest Anim = Zuchthygiene; 2012:47:293\u0026ndash;8.\u003c/li\u003e\n \u003cli\u003eZobel R, Tkalčić S (2013) Efficacy of ozone and other treatment modalities for retained placenta in dairy cows. Reprod Domest Anim = Zuchthygiene 2013:48:121\u0026ndash;5.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"veterinary-research-communications","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"verc","sideBox":"Learn more about [Veterinary Research Communications](https://www.springer.com/journal/11259)","snPcode":"11259","submissionUrl":"https://submission.nature.com/new-submission/11259/3","title":"Veterinary Research Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Haematology, Liver enzymes, Ozone therapy, Transrectal, Sheep","lastPublishedDoi":"10.21203/rs.3.rs-8405432/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8405432/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The administration of ozone has been shown to benefit human and animal health in various areas because of its anti-inflammatory, antibacterial, antiparasitic, and antiviral effects. The effects of ozone on the health status of farm animals have not yet been described. To evaluate the clinical, haematological, and biochemical responses of sheep following rectal administration of an oxygen–ozone mixture, 10 sheep were used and divided into two experimental groups: the oxygen group (GO2), which was subjected to rectal insufflation with oxygen, and the ozone group (GO2/O3), which was subjected to rectal insufflation with an oxygen-ozone mixture. The treatments were carried out three times a week for four weeks. The animals were clinically assessed before and after all applications, and blood samples were taken at five time points. The data were analysed via analysis of variance and Tukey's test (p\u003c0.05). There were significant differences between the groups in the number of total leucocytes and the serum concentrations of GGT, total protein, albumin, and bilirubin. The administration of the oxygen-ozone mixture had no significant effect on heart rate, red blood cell count, globular volume, haemoglobin concentration, or serum AST activity. The results showed that treatment with the oxygen-ozone mixture did not lead to significant clinical and/or haematological changes or deleterious liver changes in sheep. Therefore, ozone therapy should continue to be studied as an alternative treatment option in sheep.","manuscriptTitle":"Clinical, haematological, and biochemical responses of rectal oxygen-ozone administration in sheep","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-25 13:36:04","doi":"10.21203/rs.3.rs-8405432/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-01-08T15:40:11+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-07T09:20:51+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-04T20:19:47+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-03T10:26:50+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-02T21:39:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"313723042908882147556058911933127431865","date":"2025-12-29T08:37:15+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"140758679407152689975402571634511298555","date":"2025-12-25T12:34:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"67521777937456824670659480068922263577","date":"2025-12-25T12:01:19+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"300159685557162031179646025125277626572","date":"2025-12-23T13:48:12+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"77054555279778759293091098385379771488","date":"2025-12-23T13:25:29+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"23076763625254712196542043914753801491","date":"2025-12-23T13:16:14+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-12-23T11:51:10+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-12-23T09:11:30+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-12-23T09:11:07+00:00","index":"","fulltext":""},{"type":"submitted","content":"Veterinary Research Communications","date":"2025-12-19T13:32:53+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"veterinary-research-communications","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"verc","sideBox":"Learn more about [Veterinary Research Communications](https://www.springer.com/journal/11259)","snPcode":"11259","submissionUrl":"https://submission.nature.com/new-submission/11259/3","title":"Veterinary Research Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"955a5801-4a50-47e0-9207-29b586f2c77d","owner":[],"postedDate":"December 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-04-20T16:07:02+00:00","versionOfRecord":{"articleIdentity":"rs-8405432","link":"https://doi.org/10.1007/s11259-026-11205-4","journal":{"identity":"veterinary-research-communications","isVorOnly":false,"title":"Veterinary Research Communications"},"publishedOn":"2026-04-13 15:59:33","publishedOnDateReadable":"April 13th, 2026"},"versionCreatedAt":"2025-12-25 13:36:04","video":"","vorDoi":"10.1007/s11259-026-11205-4","vorDoiUrl":"https://doi.org/10.1007/s11259-026-11205-4","workflowStages":[]},"version":"v1","identity":"rs-8405432","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8405432","identity":"rs-8405432","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}