Research Article | 14 Oct 2025

In vitro antibacterial activity of Piper betel extract nanoemulsion and cannabidiol formulations against methicillin-resistant Staphylococcus isolates from canine pyoderma

Wongsakorn Wongwatcharamongkhon1 , Pareeya Udomkusonsri2 , Natthasit Tansakul2 , Udomlak Sukatta3 , Suporn Thongyuan4 , Watsapol Suntoranan1 , and Chantima Pruksakorn1 Show more
VETERINARY WORLD | pg no. 3017-3028 | Vol. 18, Issue 10 | DOI: 10.14202/vetworld.2025.3017-3028
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Abstract

          
Background and Aim: Canine pyoderma is a common dermatological condition, often caused by Staphylococcus pseud­intermedius and related methicillin-resistant strains (MRSP and MRSS). Rising antimicrobial resistance necessitates alter­native topical therapies. This study comparatively evaluated the in vitro antibacterial activity of P. betel leaf extract (both solvent-based and nanoemulsion forms) and cannabidiol (CBD) formulations against canine Staphylococcus isolates.
Materials and Methods: Antibacterial activity was determined by broth microdilution to establish minimum inhibitory con­centration (MIC) and minimum bactericidal concentration (MBC). Five formulations were tested: Ethanolic betel leaf extract in dimethyl sulfoxide betel leaf (BL), BL extract nanoemulsion (BLN), CBD in ethanol, water-soluble CBD, and CBD nanoemul­sion. Test organisms included 15 Staphylococcus isolates (five MRSP, five methicillin-susceptible S. pseudintermedius, and five MRSS) and five Pseudomonas aeruginosa. Gas chromatography-mass spectrometry (GC-MS) was used to characterize phytochemical constituents.
Results: GC-MS revealed eugenol (40.86%) and hydroxychavicol (26.44%) as predominant antibacterial compounds. BL and BLN demonstrated potent anti-staphylococcal activity, with median MICs of 0.16 g/L and 0.31 g/L, respectively. BL exhibited significantly lower MIC and MBC values than BLN (p = 0.008). Among CBD formulations, ethanol-dissolved and water-sol­uble CBD displayed the strongest activity (median MICs 0.003 g/L and 0.004 g/L), while CBD nanoemulsion was markedly less effective (median MIC 7.50 g/L). BLN also exhibited antibacterial activity against P. aeruginosa (median MIC 0.62 g/L), comparable to BL.
Conclusion: The novel BLN and soluble CBD formulations demonstrated significant in vitro antibacterial activity against methicillin-resistant Staphylococcus isolates from canine pyoderma. These results highlight their potential as topical anti­septic alternatives to chlorhexidine. Further in vivo studies are required to assess safety, efficacy, and formulation optimiza­tion. A combined betel-CBD nanoemulsion represents a promising direction for developing novel veterinary dermatological therapies.
          
Keywords: antimicrobial resistance, canine pyoderma, cannabidiol, nanoemulsion, Piper betel, Staphylococcus pseudintermedius.