Piper betle extract and its application in bovine teat dipping solution inhibit and eliminate biofilms in bovine mastitis-inducing staphylococci

abstract

Background and Aim: Staphylococci, including Staphylococcus aureus, Staphylococcus chromogenes, and Staphylococcus haemolyticus, are significant bacteria that induce bovine mastitis, primarily because they can form biofilms in bovine teat canals. This study aimed to investigate the efficacy of Piper betle extract and a bovine teat dipping solution containing P. betle extract (BSP) against these mastitis-causing staphylococci.Materials and Methods: BSP was prepared using P. betle extract as the bioactive compound. The antibacterial activity of the plant extract and BSP against the pathogens was investigated using a broth microdilution method. The activity of the extract and BSP against the pathogen biofilms was also determined. A stability test was performed to observe the pH, color, turbidity, homogeneity, precipitation, and separation of BSP stored at 4degree celsius and 25degree celsius for up to 4 weeks.Results: The extract exhibited potent antibacterial activity against S. aureus and S. haemolyticus, with similar values for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) ranging from 0.03 mg/mL to 0.125 mg/mL. The MIC and MBC values of the extract against S. chromogenes were 0.5-1 mg/mL and 0.5-2 mg/mL, respectively. Moreover, BSP exhibited MIC and MBC values of 12.5-50 v/v against all tested staphylococci isolates. When used at 1/2 and 1/4 x MIC, the extract and BSP significantly inhibited the formation of staphylococcal biofilms (p < 0.05) in the tested strains. The results indicated that treatment with 1/2 x MIC of the extract and BSP resulted in biofilm inhibition ranging from 30%-66% and 19%-39%, respectively. Furthermore, the extract at 16 x MIC eliminated 54%-86% of established mature isolate biofilms, whereas BSP removed 41%-61% of mature biofilm viability. Storage of BSP at 4degree celsius did not change the factors associated with stability from the 1(st) to 4(th) week.Conclusion: These findings suggest that BSP may exhibit potential medicinal benefits in inhibiting the growth and biofilm formation of mastitis-inducing staphylococci in bovines.

subject category

Agriculture; Veterinary Sciences

authors

Sungkatavat, P; Khongkhai, H; Kanchana, W; Saengsawarng, P; Sangkanu, S; Nissapatorn, V; Pereira, MD; Ontong, JC; Mitsuwan, W

Groups

G5 - Biomimetic, Biological and Living Materials

Projects

CICECO - Aveiro Institute of Materials (UIDB/50011/2020)

CICECO - Aveiro Institute of Materials (UIDP/50011/2020)

Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)

acknowledgements

This work was funded by Center of Excellence in Innovation of Essential Oil and Bioactive Compounds (Grant no. WU-COE-65-05) , Walailak University, Thailand and the Project CICECO-Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 and LA/P/0006/2020, financed by national funds through the FCT/MEC (PIDDAC) . We would like to thank One Health Research Center, Walailak University, Thailand. We are also grateful to Akkhraratchakumari Veterinary College, Walailak University, Thailand.