Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis

Murat Can Demir; Mushap Kuru; Mustafa Makav; Merve Sena Demir; Semra Kaya

doi:10.47748/tjvr.1866616

Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis

Abstract

Objective: This study aimed to evaluate alterations in oxidative stress markers and thiol–disulfide homeostasis in cows with clinical and subclinical mastitis. Materials and Methods: A total of 40 cows were included and divided into five groups: clinical mastitis (n = 8), subclinical mastitis (+1, +2, +3; n = 8 for each group), and a healthy control group (n = 8). Blood samples were analyzed for glutathione (GSH), malondialdehyde (MDA), total thiol (TT), native thiol (NT), disulfide levels, and the ratios of disulfide/TT, disulfide/NT, and NT/TT. Results: Compared with the control group, cows with clinical and subclinical mastitis showed significantly lower GSH, TT, and NT levels (P < 0.001), whereas MDA, disulfide concentrations, and disulfide-based ratios were markedly increased, particularly in the clinical mastitis group (P < 0.001). Progressive alterations in oxidative stress markers were observed with increasing severity of subclinical mastitis (P < 0.05). Correlation analyses revealed strong and significant associations between thiol–disulfide homeostasis parameters and oxidative stress markers (P < 0.001). Conclusion: In conclusion, thiol–disulfide homeostasis may serve as a complementary oxidative stress–based biomarker system for the evaluation of both clinical and subclinical mastitis.

Keywords

Ethical Statement

This study was approved by the Kafkas University Local Ethics Committee for Animal Experiments (KAU-HADYEK/2025-04).

References

Abdel-Hamied E, Mahmoud MM. Antioxidants profile, oxidative stress status, leukogram and selected biochemical indicators in dairy cows affected with mastitis. J Anim Health Prod. 2020; 8(4):183-188.
Akpınar RK, Çenesiz S, Şahin B, et al. Investigation of oxidative stress parameters in Brucella infected sheep and goats. Anim Health Prod Hyg. 2022; 14(1):1-9.
Akyüz B, Kara E. Investigation of thiol disulphide balance in dogs with canine leishmaniasis. Turkish Journal of Veterinary Internal Medicine. 2025; 4(1):10-17.
Atalay MS, Dilbaz B, Kinay T, et al. Evaluation of oxidative stress with “Dynamic Thiol/Disulfide Homeostasis” in cases with endometrioma. Med Sci Discov. 2022; 9:458-464.
Atroshi F, Parantainen J, Sankari S, Österman T. Prostaglandins and glutathione peroxidase in bovine mastitis. Res Vet Sci. 1986; 40(3):361-366.
Ayala A, Muñoz MF, Argüelles S. Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxid Med Cell Longev. 2014; 2014:360438.
Balkan BM, Meral O, Cetintav B, Ozkurt G, Balkan AB, Sel T. Evaluation of thiol/disulphide and oxidant–antioxidant status of dairy cows in periparturient and post-partum period. Vet Med Sci. 2024; 10(6):e70023.
Beutler E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med. 1963; 61:882-888.

Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative stress and antioxidant defense. World Allergy Organ J. 2012; 5(1):9-19.
Boulanger V, Zhao X, Lacasse P. Protective effect of melatonin and catalase in bovine neutrophil-induced model of mammary cell damage. J Dairy Sci. 2002; 85(3):562-569.
Catalá A. A synopsis of the process of lipid peroxidation since the discovery of the essential fatty acids. Biochem Biophys Res Commun. 2010; 399(3):318-323.
Celi P. Biomarkers of oxidative stress in ruminant medicine. Immunopharmacol Immunotoxicol. 2011; 33(2):233-240.
Çamkerten İ, Çamkerten G, Erdoğan H, Ayan A, Erdoğan S, Ural K. Serum thiol disulphide levels among sheep with sarcoptic mange. Kafkas Univ Vet Fak Derg. 2019; 25(6):865-868.
Değirmençay Ş, Çamkerten G, Çamkerten İ, Aktaş MS. An investigation of thiol/disulfide homeostasis and ischemia-modified albumin levels to assess the oxidative stress in dogs with canine distemper. Vet Arhiv. 2021; 91(1):39-49.
Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis. 2005; 15(4):316-328.
Deveci H, Güven A. Investigation of blood MDA and GSH levels in cows with mastitis. Kafkas Univ Vet Fak Derg. 2008; 14(1):63-66.
Deveci MZY, Erdal H. Determination of dynamic thiol-disulfide levels in dairy cattle with foot disease. Vet. Arh. 2022; 92(6):657-666.
Erel O, Neselioglu S. A novel and automated assay for thiol/disulphide homeostasis. Clin Biochem. 2014; 47(18):326-332.
Erişir M, Kandemir FM, Yüksel M. The effect of subclinical mastitis on MDA, GSH levels and GSH-Px, CAT activities in milk of cows. Fırat Univ Vet J Health Sci. 2011; 25(2):67-70.
Ertaş F, Kızıltepe Ş, Merhan O. Investigation of Dynamic Thiol Disulfide Homeostasis in Young Cattle With Pneumonia. MAS JAPS. 2023; 8:949–954.
Gu B, Zhu Y, Zhu W, Miao J, Deng YE, Zou S. Retinoid protects rats against neutrophil-induced oxidative stress in acute experimental mastitis. Int Immunopharmacol. 2009; 9(2):223-229.
Huma ZI, Sharma N, Kour S, Tandon S, Guttula PK, Gupta MK. Putative biomarkers for early detection of mastitis in cattle. Anim Prod Sci. 2020; 60(14):1721-1736.
Jhambh R, Dimri U, Gupta VK, Rathore R. Blood antioxidant profile and lipid peroxides in dairy cows with clinical mastitis. Vet World. 2013; 6(5):271-273.
Jones DP, Liang Y. Measuring the poise of thiol/disulfide couples in vivo. Free Radic Biol Med. 2009; 47(10):1329-1338.
Kaya İ, Güven A. Determination of blood vitamin A, beta carotene and vitamin E levels in cows with mastitis. Kafkas Univ Vet Fak Derg. 2008; 14(1):57-61.
Kayar T, Ozkurt G, Erzurum O, Er K, Buyukgungor B, Gecgel BN, Karaburc MN. Thiol/Disulphide Homeostasis in the Relationship Between Body Condition Score and Oxidative Stress in Periparturient Period Holstein Heifers. Vet Med Sci. 2025; 11(3):70326.
Konvičná J, Vargová M, Paulíková I, Kováč G, Kostecká Z. Oxidative stress and antioxidant status in dairy cows during prepartal and postpartal periods. Acta Vet Brno. 2015; 84:133-140.
Kundi H, Ates I, Kiziltunc E, et al. A novel oxidative stress marker in acute myocardial infarction; thiol/disulphide homeostasis. Am J Emerg Med. 2015; 33(11):1567-1571.
Kurtdede E, Börkü MK, Gür B, Doğan Y, Sevim K, Gülendağ E. Evaluation of CBC, lipid profile, oxidative stress biomarkers, total thiol, native thiol, and disulfide levels in dogs diagnosed with parvoviral enteritis without clinical findings. Vet Sci Pract. 2022; 17(3):98-102.
Merhan O, Başer L, Bozukluhan K, Kükürt A, Lehimcioğlu NC, Atakişi E, Çelik Ö. Determination of dynamic thiol disulfide homeostasis in cows with dystocia. Animal Health Production and Hygiene. 2025; 14(2): 40-45.
Musal B, Ulutas PA, Turkyilmaz S. Blood vitamin C, vitamin A, β-carotene, ceruloplasmin, glutathione and malondialdehyde concentrations in cows with sub-clinical mastitis treated with intramammary antibiotics. Rev Med Vet. 2007; 158(12):633-640.
Nakov D, Kuzelov A, Hristov S, Nakova VV, Stanković B, Miočinović J. The impact of mastitis pathogens on antioxidant enzyme activity in cows’ milk. Contemp Agric. 2023; 72(4):199-206.
Nassif MN, Ibrahim HM, Moharam HA. Antioxidant trace elements and oxidative stress status in dairy cattle suffering from mastitis. Mansoura Vet Med J. 2017; 18(1):165-174.
Ponnampalam EN, Kiani A, Santhiravel S, Holman BWB, Lauridsen C, Dunshea FR. The importance of dietary antioxidants on oxidative stress, meat and milk production, and their preservative aspects in farm animals: antioxidant action, animal health, and product quality—invited review. Animals. 2022; 12(23):3279.
Prin-Mathieu C, Le Roux Y, Faure GC, Laurent F, Bene MC, Moussaoui F. Enzymatic activities of bovine peripheral blood leukocytes and milk polymorphonuclear neutrophils during intramammary inflammation caused by lipopolysaccharide. Clin Vaccine Immunol. 2002; 9(4):812-817.
Raju R, Shobha UK, Shastri BA, et al. Evaluation of thiol/disulfide homeostasis and ischemia-modified albumin in metabolic syndrome patients. Adv Redox Res. 2025; 14: 100116.
Sadat A, Farag AM, Elhanafi D, Awad A, Elmahallawy EK, Elshopakey GE. Immunological and oxidative biomarkers in bovine serum from healthy, clinical, and sub-clinical mastitis caused by Escherichia coli and Staphylococcus aureus infection. Animals. 2023; 13(5):892.
Salman S, Khol-Parisini A, Schafft H, et al. The role of dietary selenium in bovine mammary gland health and immune function. Anim Health Res Rev. 2009; 10:21-34.
Sandikci SC, Colak S, Omma A, et al. An investigation of thiol/disulfide homeostasis in patients with Behçet’s disease. Archives of Medical Science. 2020; 16(6):1353–1359.
Schalm OW and Noorlander DO. Experiments and observations leading to development of the California mastitis test. J Am Vet Med Assoc. 1957; 130(5):199-204.
Şenel Y, Terzi OS, Kara E, Erel Ö, Neşelioğlu S, Ceylan E. Alterations in serum thiol-disulfide homeostasis and ischemia-modified albumin concentrations in clinical canine parvoviral enteritis. Turk J Vet Anim Sci. 2024; 48(2):88-96.
Terzi OS, Kara E, Şenel Y, Ceylan E, Neşelioğlu S, Erel Ö. Dynamic thiol-disulphide homeostasis and ischemia modified albumin levels in neonatal calf diarrhea. Ankara Univ Vet Fak Derg. 2022; 70(1):81-86.
Turk R, Koledić M, Maćešić N, et al. The role of oxidative stress and inflammatory response in the pathogenesis of mastitis in dairy cows. Mljekarstvo. 2017; 67(2):91-101.
White LJ, Lam TJGM, Schukken YH, Green LE, Medley GF, Chappell MJ. The transmission and control of mastitis in dairy cows: a theoretical approach. Prev Vet Med. 2006; 74(1):67-83.
Yoshioka T, Kawada K, Shimada T, Mori M. Lipid peroxidation in maternal and cord blood and protective mechanism against activated-oxygen toxicity in the blood. Am J Obstet Gynecol. 1979; 135:372-376.

Details

Primary Language

English

Subjects

Veterinary Obstetrics and Gynecology

Journal Section

Research Article

Authors

Murat Can Demir ^*
0000-0002-3803-069X
Türkiye

Mushap Kuru
0000-0003-4409-251X
Türkiye

Mustafa Makav
0000-0003-1879-8180
Türkiye

Merve Sena Demir
0000-0002-2630-8818
Türkiye

Semra Kaya
0000-0002-7520-6631
Türkiye

Publication Date

March 29, 2026

Submission Date

January 19, 2026

Acceptance Date

February 16, 2026

Published in Issue

Year 2026 Volume: 10 Number: 1

DOI

https://doi.org/10.47748/tjvr.1866616

IZ

https://izlik.org/JA73RZ58FP

Cite

RIS / Bibtex

APA

Demir, M. C., Kuru, M., Makav, M., Demir, M. S., & Kaya, S. (2026). Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis. Turkish Journal of Veterinary Research, 10(1), 93-100. https://doi.org/10.47748/tjvr.1866616

AMA

1.Demir MC, Kuru M, Makav M, Demir MS, Kaya S. Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis. TJVR. 2026;10(1):93-100. doi:10.47748/tjvr.1866616

Chicago

Demir, Murat Can, Mushap Kuru, Mustafa Makav, Merve Sena Demir, and Semra Kaya. 2026. “Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows With Clinical and Subclinical Mastitis”. Turkish Journal of Veterinary Research 10 (1): 93-100. https://doi.org/10.47748/tjvr.1866616.

EndNote

Demir MC, Kuru M, Makav M, Demir MS, Kaya S (March 1, 2026) Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis. Turkish Journal of Veterinary Research 10 1 93–100.

IEEE

[1]M. C. Demir, M. Kuru, M. Makav, M. S. Demir, and S. Kaya, “Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis”, TJVR, vol. 10, no. 1, pp. 93–100, Mar. 2026, doi: 10.47748/tjvr.1866616.

ISNAD

Demir, Murat Can - Kuru, Mushap - Makav, Mustafa - Demir, Merve Sena - Kaya, Semra. “Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows With Clinical and Subclinical Mastitis”. Turkish Journal of Veterinary Research 10/1 (March 1, 2026): 93-100. https://doi.org/10.47748/tjvr.1866616.

JAMA

1.Demir MC, Kuru M, Makav M, Demir MS, Kaya S. Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis. TJVR. 2026;10:93–100.

MLA

Demir, Murat Can, et al. “Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows With Clinical and Subclinical Mastitis”. Turkish Journal of Veterinary Research, vol. 10, no. 1, Mar. 2026, pp. 93-100, doi:10.47748/tjvr.1866616.

Vancouver

1.Murat Can Demir, Mushap Kuru, Mustafa Makav, Merve Sena Demir, Semra Kaya. Alterations in Thiol–Disulfide Homeostasis and Oxidative Stress Markers in Cows with Clinical and Subclinical Mastitis. TJVR. 2026 Mar. 1;10(1):93-100. doi:10.47748/tjvr.1866616