Therapeutic potential of an ethnoveterinary formulation against Staphylococcus aureus-induced mastitis via modulating...

Mastitis is a major inflammatory disorder of the mammary gland that causes substantial economic losses in the dairy industry, with Staphylococcus aureus (S. aureus) being one of the predominant causative pathogens. The current treatment options are limited by antimicrobial resistance and drug residues. Therefore, alternative therapeutic strategies are needed. The present study aimed to evaluate the anti-inflammatory and antioxidant effects of an ethnoveterinary oil (EO) formulation in an experimental model of S. aureus-induced mastitis. The EO formulation was prepared according to a standardized traditional ethnoveterinary composition. Mastitis was induced in healthy lactating primiparous Wistar rats by intramammary infusion of S. aureus (2 × 10⁸ CFU/mL), and the twenty-four female Wistar rats were divided into four groups of 6: normal control (NC), mastitis control (MT), standard drug-treated (MT + DEX), and MT + EO-treated groups. The chemical composition of EO was characterized by GC-MS analysis, which identified 21 bioactive compounds, predominantly erucic acid, along with phenolic compounds, fatty acid derivatives, terpenoids, and steroidal constituents known for their anti-inflammatory potential. Biochemical analyses revealed that EO significantly restored endogenous antioxidant enzyme activities and reduced lipid peroxidation in mammary tissues. Furthermore, EO significantly decreased myeloperoxidase (MPO) levels and N-acetyl-β-D-glucosaminidase activity, indicating reduced neutrophil infiltration and protection against epithelial cell damage. The levels of Pro-inflammatory cytokines were markedly elevated in mastitis control group whereas EO treatment significantly suppressed their production. In addition, EO downregulated the gene expression of Toll like receptor-2 (TLR-2) and Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX-2), suggesting modulation of key inflammatory and oxidative stress-related pathways. Histopathological and special staining analyses further confirmed the preservation of mammary gland architecture and attenuation of inflammatory infiltration following EO treatment. Collectively, these findings demonstrate that EO exerts potent anti-mastitic effects through the combined modulation of oxidative stress and inflammatory responses and may serve as a promising natural therapeutic alternative for the management of S. aureus associated mastitis in Wistar rat model.