Antibacterial and antibiofilm activity of methanolic Indian propolis extract against ESBL Pseudomonas isolated from buffalo mastitis

ASHOK  BOORA; SARITA  YADAV; SUNITA  YADAV; ASHOK  BALHARA; SUNESH BALHARA; NISHU DHAKA; NISHA VERMA; ANIL  KUMAR; RAJESH KUMAR  LOHCHAB

doi:10.56093/ijans.v95i6.169361

Authors

ASHOK BOORA ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125 001, India
SARITA YADAV ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125 001, India
SUNITA YADAV Chaudhary Charan Singh Haryana Agricultural University, Hisar 125 001
ASHOK BALHARA ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125 001, India
SUNESH BALHARA ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125 001, India
NISHU DHAKA ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125 001, India
NISHA VERMA ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125 001, India
ANIL KUMAR Indian Council of Agricultural Research, New Delhi 110 001, India
RAJESH KUMAR LOHCHAB Guru Jambheshwar University of Science & Technology, Hisar 125 001

https://doi.org/10.56093/ijans.v95i6.169361

Keywords:

Antibacterial, Antibiofilm, Environmental mastitis ESBL, Propolis, Pseudomonas, Minimum inhibitory concentration

Abstract

Increasing antimicrobial resistance poses a major challenge to mastitis treatment in dairy animals, highlighting the need for alternative therapeutics. This study analysed the antibacterial and antibiofilm activity of methanolic propolis extract (MPE) against extended-spectrum β-lactamase (ESBL) positive Pseudomonas spp. recovered from mastitis affected Murrah buffaloes. Of the 175 environmental isolates from mastitis-affected buffalo milk (n= 472 animals), Pseudomonas spp. accounted for 23.4% (n=41), making it the second most prevalent pathogen after Escherichia coli (24.6%). Six clinical ESBL positive Pseudomonas isolates were selected for analysis. MPE exhibited antibacterial activity with minimum inhibitory concentration (MIC) in the range of 7.81 to 15.62 mg/mL. Scanning electron microscopy and fluorescence microscopy revealed membrane disruption in treated cells, and flow cytometry further confirmed a high proportion of apoptotic cells post-treatment. All isolates were strong biofilm producers, and MPE showed significant antibiofilm activity with 72.5± 0.39% and 71.2 ± 0.95% biofilm inhibition at 2MIC and MIC, respectively. The findings indicate that Indian propolis is a promising natural alternative with potent antibacterial and antibiofilm activity against Pseudomonas associated with bovine mastitis.

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