Alicyclobacillus in Orange Juice: Occurrence and Heat Resistance of Spores

Abstract

Spore suspensions of a pure culture of Alicyclobacillus acidoterrestris DSM 2498 were submitted to different heat treatments (60°C for 60 min, 60°C for 30 min, 70°C for 20 min, 80°C for 5 min, 80°C for 10 min, 80°C for 30 min, and boiling for 5 min) to determine the best activation conditions in orange juice. The best treatment for spore activation was shown to be 70°C/20 min. Seventy-five samples of concentrated orange juice from 11 different suppliers were examined for the presence of thermophilic acid-tolerant spore formers by the most probable number technique using Bacillus acidocaldarius medium (BAM broth) and incubation at 44°C for 5 days after a prior spore activation. After incubation, isolation was carried out using BAM agar medium incubating at 44°C for 5 days. Typical colonies were submitted to a microscopic examination, evaluation for the presence of spores, and various biochemical tests. Of the orange juice samples examined, 14.7% were found to be positive for Alicyclobacillus. The thermal death time open tube method was used to determine the heat resistance of the spores of strains confirmed as being Alicyclobacillus. The D-values determined were in the range from 60.8 to 94.5 min at 85°C, 10.0 to 20.6 min at 90°C, and 2.5 to 8.7 min at 95°C. The z-values were between 7.2°C and 11.3°C. The results demonstrated the occurrence of Alicyclobacillus in orange juice and the high heat resistance of the spores that could survive the heat treatments normally applied in the processing of orange juice.