Abstract

Antibiotic therapy has been the most popular line of treatment for the control of mastitis worldwide during the last few decades. Alternative and sustainable treatments must be developed because pathogens are becoming more resistant to antibiotics, leading to the development and spread of antimicrobial resistance (AMR). The aim of the current investigation was to isolate lactic acid bacteria (LAB) with probiotic potential that can inhibit mastitis-causing pathogens to prevent bovine mastitis. Milk samples were collected from Sahiwal cows, and a total of 150 bacteria were isolated, of which 76 were found to be catalase negative, and resistant to vancomycin. Twenty-three isolates displayed greater acid and bile tolerance, with > 90% survivability, and were molecularly characterized by 16S rRNA partial sequencing. The autoaggregation percentages for SML7 and SML41 were greater (p<0·05) 80.38±0.19% and 80.28±0.04%, respectively. SML10 (92.04±0.26 μmol/mL) had the highest (p<0.05) ferric-reducing antioxidant power (FRAP) activity, while SML20 (52.1±0.99%) had the highest 1,1 diphenyl 2 picrylhydrazyl (DPPH) scavenging activity. All the strains were nonhemolytic or nonmucinolytic. The highest antimicrobial activity was observed in several strains (SML41, SML63, SML76, and SML60) against common mastitis-causing pathogens, namely, E. coli ATCC25922, Staphylococcus aureus ATCC25923, Enterococcus faecalis NCDC114, Streptococcus agalactiae NCDC208, and Enterococcus faecium NCDC124. The coaggregation efficacy of SML20 with S. aureus was the highest (67.69±1.21%), while SML41 showed the highest (69.75±0.29%) coaggregation efficacy with E. faecalis NCDC114 and SML63 (68.078±0.26) with S. agalactiae NCDC208. Overall, seven distinct lactic acid bacterial clusters were identified by cluster analysis of the phylogenetic tree as follows: Enterococcus hirae (1), Limosilactobacillus reuteri (1), Pediococcus acidilactici (4), Weissella confusa (11), Lactobacillus helveticus (3), Limosilactobacillus balticus (2), and Lacticaseibacillus rhamnosus (1). The Lactobacillus helveticus SML41, Lactobacillus helveticus SML60, Weissella confusa SML61, Lacticaseibacillus rhamnosus SML63, Weissella confusa SML64, and Pediococcus acidilactici SML76 isolates were found to possess the most desirable characteristics of potential probiotics based on principal component analysis (PCA). Therefore, the strains chosen in the current investigation demonstrated techno-functional characteristics that rendered them appropriate for probiotic use to treat and prevent intramammary infections in dairy cattle in a sustainable manner.

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