In vitro anti-biofilm effects of Loxostylis alata extracts and isolated 5-demethyl sinensetin on selected foodborne bacteria

Abstract

Bacterial biofilms pose health challenges both in clinical environments and the food industry. Major foodborne bacterial pathogens form biofilms on surfaces and persist, causing infections in humans that may be difficult to treat. Conventional use of antibiotics is fast becoming ineffective due to emerging resistance of pathogens to antibiotics. Previous studies have demonstrated the antimicrobial potential of Loxostylis alata A. Spreng. ex Rchb. extracts against a range of bacterial pathogens. The inhibitory effects of methanol and aqueous extracts of L. alata and an antibacterial compound (5-demethyl sinensetin) isolated from the leaves on the growth and development of microbial biofilms was investigated against clinical isolates of Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis and reference strains of Bacillus cereus, Salmonella Enteritidis and Salmonella Typhimurium using the crystal violet (CV) assay. The Kirby-Bauer disk diffusion method was utilized to determine antibiotic susceptibility and resistance profiles of the four clinical and three reference isolates. E. cloacae was resistant to tetracycline and doxycycline and P. mirabilis was resistant to colistin while E. coli and K. pneumoniae were pan-susceptible to all the antibiotics screened. The hot water extract reduced biofilm adhesion from 7% to >50% for the clinical isolates and the ATCC strains, except for S. Typhimurium. The methanol extract inhibited the growth of preformed biofilms of S. Enteritidis and S. Typhimurium by >50% and also reduced the biofilm biomass in S. Enteritidis and E. coli by >50%. The plant extracts and compound were able to reduce initial cell attachment and biofilm biomass, although inhibition of growth in a preformed biofilm was not attained.