the apple juice samples.The standard method of FDA’s Bacteriol ogical
Analytical Manual Methods [16] was used to determine the
microbial load in the juice sample.To determine the total plate
counts in the samples, pour plate method was used.Proper serial
dilution was made by using sterilized distilled water followed by
further decimal dilution of sample (upto 10 5) which was then
poured into sterile petri dishes by using pipette.To each petri dish
15 mL of molten agar was added.Then immediatel y sample dilu-
tions were mixed with agar medium by moving each petri dish five
times in a vertical,clockwise,horizontal and anticlockwi se direction
and then allowed the dishes to set for 30 min at 25 ±1 C.
The plates were turned upside down and placed in an incubato r
(GSP-9080 MBE,Shanghai Boxun Industry &Commerce Co.,Ltd,
China) at 32 C for 2days.The numbers of bacterial colonies in
sample (as CFU/mL juice)were counted by multiplyi ng with reciprocal
and the results were mentioned as log colony-forming units
(CFU)/mL of juice.
PDA media was used to determine the total yeast and mold
counts by pour plate method. Media was prepared by dissolving
a known amount of PDA powder (39g)in distilled water
(1000 mL).To protect the PDA agar from the growth of other mi-
crobes by cross contamination , tartaric acid (10%)was added.All
the PDA plates were placed in an incubator at 32 ±1 C. Yeast
and mold counts were determined in each plate after 2days of
incubation and the results were expressed as log colony-forming
units (CFU)/mL of juice.All the analyses were performed in
triplicate .