Time-kill kinetics and biocidal effect of Euclea crispa leaf extracts against microbial membrane.

Abstract

To evaluate antimicrobial potential of the fractions partitioned from Euclea crispa leaf extract and determination of their impact on cell membrane disruption. Antimicrobial potentials were evaluated via susceptibility test, determination of minimum inhibitory concentrations (MICs) and time-kill kinetics of the potent fractions. Degree of membrane disruption was determined by the amount of proteins and nucleotides released from within the cells and SEM images of the membrane after 120min of treatment. The largest inhibition zone (25.5±0.50mm) was obtained by ethylacetate fraction against Aeromonas hydrophilla at 10mg/mL. The lowest MIC (0.16mg/mL) was exhibited by n-butanol and ethylacetate fractions against test bacteria while all fractions exhibited MIC values between 0.31 and 1.25mg/mL against susceptible yeast. n-Butanol fraction achieved absolute mortality against Bacillus pumulis (B.pumulis) and Klebsiella pneumoniae (K.pneumoniae) after 90 and 120min contact time respectively at 1×MIC. Total mortality also achieved by n-hexane fraction against B.pumulis and K.pneumoniae after 90 and 120min respectively at 2×MIC. Ethylacetate fraction achieved absolute mortality against both bacteria after 120min at 2×MIC. n-Hexane fraction achieved total mortality against Candida albicans after 120min at 1×MIC. Maximum amount of proteins (0.566μg/mL) was released from K.pneumoniae by n-butanol fraction at 2×MIC after 120min of treatment while the maximum amount of nucleotides released (4.575μg) was from B.pumulis by n-hexane fraction under similar condition. This study suggests the leaf of Euclea crispa a source of bioactive compound with membrane attack as one of the mechanisms of its biocidal action.