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Animal reproduction2026; 23(3); e20250116; doi: 10.1590/1984-3143-AR2025-0116

Antibiofilm action of ozonated sunflower oil against bacteria isolated from the uterus of mares susceptible to endometritis.

Abstract: This study aimed to characterize the antibiofilm action of ozone-enriched sunflower oil (O) against pathogens associated with endometritis in mares. Initially, samples from 41 mares with a history of subfertility were stored in BHI broth and 0.9% NaCl solution. The samples underwent microbial culture, identification, and sensitivity testing at LABRAPE-UFRPE, Brazil. Biofilm production was evaluated by the crystal violet method. Subsequently, biofilm-forming bacteria were subjected to the action of different concentrations of ozonated sunflower oil. Of the 21 isolates tested for biofilm production, 76.2% (16/21) showed positive production, with 43.75% (7/16) being weak producers, 31.25% (5/16) moderate producers, and 25% (4/16) strong producers. After the action of ozonated sunflower oil at a concentration of 203.43 mEq/L, a reduction was observed in the mass of the gram-negative bacilli biofilm, however, no statistical difference was found for the gram-positive cocci biofilm. The results show that ozonated sunflower oil presented satisfactory results in breaking down gram-negative bacteria biofilm.
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Publication Date: 2026-07-06 PubMed ID: 42524657PubMed Central: PMC13412003DOI: 10.1590/1984-3143-AR2025-0116Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Objective Overview

  • This research investigated the effectiveness of ozonated sunflower oil in disrupting biofilms formed by bacteria isolated from the uterus of mares prone to endometritis.
  • The study particularly focused on how this oil affects biofilm production by different bacterial isolates derived from mares with subfertility issues associated with uterine infections.

Introduction and Background

  • Endometritis is a common uterine infection in mares that can lead to subfertility and reproductive problems.
  • Bacteria involved in endometritis often form biofilms, which are structured communities of microorganisms attached to surfaces enclosed in a protective matrix.
  • Biofilms contribute to bacterial resistance against treatment and the immune system, making infections difficult to eradicate.
  • Ozonated sunflower oil is enriched with ozone, known for its antimicrobial properties, and is being explored as a treatment to disrupt biofilms and improve infection control.

Methods

  • Samples were collected from 41 mares with a history of subfertility and likely uterine infections.
  • These samples were stored in brain heart infusion (BHI) broth and saline (0.9% NaCl) to maintain bacterial viability until testing.
  • Microbial cultures were grown, and bacteria were identified along with sensitivity testing at LABRAPE-UFRPE, Brazil.
  • Biofilm production ability of the isolates was evaluated using the crystal violet assay, which stains biofilm biomass for quantification.
  • Bacteria that produced biofilms were then exposed to varying concentrations of ozonated sunflower oil to assess its antibiofilm action.

Results

  • Out of 21 isolates tested for biofilm formation, 76.2% (16 isolates) were positive biofilm producers.
  • Among these, 43.75% (7 isolates) were weak biofilm producers, 31.25% (5 isolates) were moderate producers, and 25% (4 isolates) were strong producers.
  • When exposed to ozonated sunflower oil at 203.43 mEq/L, a significant reduction in biofilm mass was observed specifically in gram-negative bacilli.
  • However, no statistically significant reduction in biofilm mass was observed for gram-positive cocci following treatment.

Conclusions and Implications

  • Ozonated sunflower oil demonstrated effective antibiofilm activity against gram-negative bacterial biofilms isolated from the uterus of mares.
  • This suggests potential use of ozonated sunflower oil as a therapeutic option to disrupt biofilms related to uterine infections, particularly those caused by gram-negative bacteria.
  • The lack of significant effect on gram-positive cocci biofilms indicates that the oil’s antibiofilm efficacy may be selective and may require complementary treatments for mixed infections.
  • Further research could explore optimization of concentration, exposure time, and combination therapies to enhance broad-spectrum antibiofilm activity in veterinary reproductive health.

Cite This Article

APA
da Silva AB, Freire LQB, Leite MEP, Martins LR, de Sobral GG, Mendonça M, da Silva ER, Leal HV, Carneiro GF. (2026). Antibiofilm action of ozonated sunflower oil against bacteria isolated from the uterus of mares susceptible to endometritis. Anim Reprod, 23(3), e20250116. https://doi.org/10.1590/1984-3143-AR2025-0116

Publication

Animal reproduction
ISSN: 1984-3143
NlmUniqueID: 101272804
Country: Brazil
Language: English
Volume: 23
Issue: 3
Pages: e20250116
PII: e20250116

Researcher Affiliations

da Silva, Antonio Brito
  • Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco - UFRPE, Recife, PE, Brasil.
Freire, Laís Querino Barbosa
  • Laboratório de Reprodução Animal de Pernambuco, Universidade Federal do Agreste de Pernambuco - UFAPE, Garanhuns, PE, Brasil.
Leite, Maria Eduarda Pedrosa
  • Laboratório de Reprodução Animal de Pernambuco, Universidade Federal do Agreste de Pernambuco - UFAPE, Garanhuns, PE, Brasil.
Martins, Lamartine Rodrigues
  • Universidade Federal de Pernambuco - UFPE, Recife, PE, Brasil.
de Sobral, Gilvannya Gonçalves
  • Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco - UFRPE, Recife, PE, Brasil.
Mendonça, Marcelo
  • Laboratório de Microbiologia e Imunologia, Universidade Federal do Agreste de Pernambuco - UFAPE, Garanhuns, PE, Brasil.
da Silva, Elizabete Rodrigues
  • Laboratório de Microbiologia, Universidade Federal do Agreste de Pernambuco - UFAPE, Garanhuns, PE, Brasil.
Leal, Haroldo Vargas
  • Horses Vet. Services, Juiz de Fora, MG, Brasil.
Carneiro, Gustavo Ferrer
  • Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco - UFRPE, Recife, PE, Brasil.

Conflict of Interest Statement

Conflicts of interest: The authors have no conflict of interest to declare.

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