The pellets were washed by sterile buffered peptone water then resuspended in 500 mL of sterile orange juice aliquots to inoculate 29.5 L orange juice before packaging.
Samples were taken from final aliquot, plated on MRS agar and incubated for 48 h at 37 °C under CO2 atmosphere (5%) to determine initial microbial population.
Packages were prepared by a hand heat sealer using antimicrobial nanocomposite and pure LDPE films 15 × 10 cm in size, similar to Doypack packaging commonly used for packaging fruit juice.
The packages were immediately wrapped in aluminum foil and sanitized at 95 °C for 2 min.
After cooling and under a sterile laboratory hood, 175 mL of inoculated orange juice was poured into each package and sealed by the heat sealer.