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Effect of uterine ozone therapy and anticoagulant sampling on oxidative stress parameters in mares

Grant number: 20/03024-1
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): May 01, 2020
Effective date (End): December 31, 2020
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Pathology
Principal Investigator:Breno Fernando Martins de Almeida
Grantee:Lidiana Zanetti Amatti
Home Institution: Faculdades Integradas de Ourinhos (FIO). Fundação Educacional Miguel Mofarrej. Ourinhos , SP, Brazil

Abstract

Ozone therapy acts in the body in order to cause a controlled oxidative stress, inducing an improvement in the antioxidant, immune and circulatory responses, which seems to assist in infectious and inflammatory processes, according to recent scientific literature. However, as ozone therapy would affect the parameters indicating oxidative stress, these could be monitored in order to obtain the best antioxidant performance without causing damage to the body. Bearing in mind that factors such as dose, time after application and number of applications are not fully standardized, especially in horses, this type of evaluation is necessary to obtain the best therapeutic effects of ozone therapy in the body. In this context, the present project will aim, firstly, to assess systemic oxidative stress in mares submitted to uterine ozone therapy, as well as to determine the best method for assessing the total antioxidant capacity (TAC); second, to assess the effect of anticoagulants on markers of oxidative stress in mares before and after the induction of oxidative stress by ozone therapy. For this purpose, 10 clinically healthy mares will be selected and will undergo the uterine ozone therapy protocol by uterine lavage with ozonated solution followed by uterine insufflation with the oxygen-ozone gas mixture for three consecutive days, with blood samples collected before (baseline), on the third , sixth, tenth and seventeenth days. Biochemical analyzes of oxidative stress will be performed in a photocolorimeter, with CAT being determined by four methods: by reducing the ABTS cation (TAC-ABTS), by inhibiting the ABTS cation associated with peroxidase (TAC-ABTS + HRP), by the reduction capacity plasma plasma (TAC-FRAP) and the copper-reducing antioxidant capacity (TAC-CUPRAC); in addition to the determination of total oxidative capacity (TOC), lipid peroxidation by substances reactive to thiobarbituric acid (TBARS), the antioxidants uric acid and albumin. The variables will be tested for normality and the differences between the moments will be verified by ANOVA tests with repeated measures and Tukey's or Friedmann's post-test with Dunn's post-test, while comparisons with the different types of sample will be performed with test multiple t, being considered significant when p <0.05.