The antibacterial activity of
PUM against S. pyogenes was evaluated by the micro
dilution method. Clinical samples obtained from the Voluntary Health
Services (VHS), Adyar, Chennai, and standard cultures obtained from
The Microbial Type Culture Collection and Gene Bank (MTCC) were grown
in Mueller–Hinton broth (MHB) supplemented with 5% defibrinated
sheep blood and used for the activity studies. The inoculum was prepared
from a single colony in MHB liquid medium with 5% defibrinated sheep
blood and incubated at 37 °C in a candle jar for 24–48
h. The derived bacterial suspension after 24 h was diluted to 108 CFU/mL (turbidity = McFarland barium sulfate standard 0.5)
with sterile MHB medium. PUM was dissolved in sterile water to a final
concentration of 1 mg/mL and further serially diluted in a 1:1 ratio
to the concentrations ranging from 1 to 0.0156 μg/mL; 100 μL
of each dilution was distributed in 96-well microtitration plates,
along with sterility control (MHB alone) and growth control (MTCC
culture). All the test and growth control wells were inoculated with
5 μL of bacterial suspension and the 96-well microtitration
plates were incubated for 24 h in a candle jar at 37 °C. The
experiments were performed in triplicate, and the microtitration plates
were checked for inhibition by streaking each well in the MHB agar
plate with 5% defibrinated sheep blood incubated at 37 °C for
24 h.
PUM against S. pyogenes was evaluated by the micro
dilution method. Clinical samples obtained from the Voluntary Health
Services (VHS), Adyar, Chennai, and standard cultures obtained from
The Microbial Type Culture Collection and Gene Bank (MTCC) were grown
in Mueller–Hinton broth (MHB) supplemented with 5% defibrinated
sheep blood and used for the activity studies. The inoculum was prepared
from a single colony in MHB liquid medium with 5% defibrinated sheep
blood and incubated at 37 °C in a candle jar for 24–48
h. The derived bacterial suspension after 24 h was diluted to 108 CFU/mL (turbidity = McFarland barium sulfate standard 0.5)
with sterile MHB medium. PUM was dissolved in sterile water to a final
concentration of 1 mg/mL and further serially diluted in a 1:1 ratio
to the concentrations ranging from 1 to 0.0156 μg/mL; 100 μL
of each dilution was distributed in 96-well microtitration plates,
along with sterility control (MHB alone) and growth control (MTCC
culture). All the test and growth control wells were inoculated with
5 μL of bacterial suspension and the 96-well microtitration
plates were incubated for 24 h in a candle jar at 37 °C. The
experiments were performed in triplicate, and the microtitration plates
were checked for inhibition by streaking each well in the MHB agar
plate with 5% defibrinated sheep blood incubated at 37 °C for
24 h.
