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

Research (Published online: 18-10-2023)

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

Paparwee Sungkatavat, Haemarat Khongkhai, Wilasinee Kanchana, Phirabhat Saengsawarng, Suthinee Sangkanu, Veeranoot Nissapatorn, Maria de Lourdes Pereira, Julalak Chorachoo Ontong, and Watcharapong Mitsuwan

Veterinary World, 16(10): 2135-2142

Paparwee Sungkatavat: Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand; One Health Research Center, Walailak University, Nakhon Si Thammarat, 80160, Thailand.

Haemarat Khongkhai: Division of Biological Science, Faculty of Sciences, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand.

Wilasinee Kanchana: Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand.

Phirabhat Saengsawarng: Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand; One Health Research Center, Walailak University, Nakhon Si Thammarat, 80160, Thailand.

Suthinee Sangkanu: School of Allied Health Sciences, Southeast Asia Water Team, World Union for Herbal Drug Discovery, and Research Excellence Center for Innovation and Health Products, Walailak University, Nakhon Si Thammarat, Thailand.

Veeranoot Nissapatorn: School of Allied Health Sciences, Southeast Asia Water Team, World Union for Herbal Drug Discovery, and Research Excellence Center for Innovation and Health Products, Walailak University, Nakhon Si Thammarat, Thailand.

Maria de Lourdes Pereira: CICECO-Aveiro Institute of Materials and Department of Medical Sciences, University of Aveiro, 3810-193 Aveiro, Portugal.

Julalak Chorachoo Ontong: Cosmetic Technology and Dietary Supplement Products Program, Faculty of Agro and Bio Industry, Thaksin University, Phatthalung, Thailand.

Watcharapong Mitsuwan: Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand; One Health Research Center, Walailak University, Nakhon Si Thammarat, 80160, Thailand; Center of Excellence in Innovation of Essential Oil and Bioactive Compounds, Walailak University, Nakhon Si Thammarat, 80160, Thailand.

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 4°C and 25°C 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 × 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 × MIC of the extract and BSP resulted in biofilm inhibition ranging from 30%–66% and 19%–39%, respectively. Furthermore, the extract at 16 × MIC eliminated 54%-86% of established mature isolate biofilms, whereas BSP removed 41%-61% of mature biofilm viability. Storage of BSP at 4°C did not change the factors associated with stability from the 1st to 4th 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.

Keywords: antibacterial activity, biofilms, bovine teat dipping solution, Piper betle extract, staphylococci.