Evaluation of bacteriological techniques, sensory evaluation, gas chromatography, and electronic nose technology for the early detection of Alicyclobacillus acidoterrestris in fruit juices

Abstract

Alicyclobacillus acidoterrestris is a spore-forming spoilage micro-organism found in fruit juices whose spores are not destroyed by typical pasteurisation. Once its spores outgrow and multiply in finished juice products, they produce two volatile taint compounds namely guaiacol and 2,6-dibromophenol. In the food industry margins for errors are small and monitoring of products to avert such errors is crucial. Conventional microbiological monitoring is one such technique for spoilage micro-organisms another being automated systems which can detect taints. Both these categories were evaluated in this study with the electronic nose and gas chromatograph being the specific automated systems being assessed. Sensory evaluation was also assessed as a diagnostic tool in the detection of taints. Isolation and identification of what was thought to be A. acidoterrestris was a laborious and expensive exercise which eventually proved inconclusive. A pure culture was purchased and juices were then inoculated with two levels of A. acidoterrestris spores and incubated. Juices from each level of inoculation were evaluated at different time intervals via the above-mentioned monitoring techniques. Of the three media assessed in the microbiological method, Bacillus acidoterrestris medium (BAM) was found to be the most effective for enumerating A. acidoterrestris followed by K-medium (KM) then Orange Serum Agar (OSA). While BAM was still indicating the presence of A. acidoterrestris KM and OSA were not (counts of <10cfu/g). This illustrated that this micro-organism could be easily overlooked if KM or OSA were being used to enumerate them. Considering that many workers actually do use KM and OSA as their media of choice in enumerating A. acidoterrestris (perhaps because BAM is very tedious to prepare) the cause for concern is a real one. Assessment of the resultant taints via sensory evaluation after inoculation and incubation reveals the inability of many panellists to detect taints at levels (as assessed by GC) far above their documented threshold values. While GC is an extremely useful and powerful tool, the level of expertise and skill required to use such an instrument cannot be overlooked nor can the expense involved. With regard to the electronic nose assessment for the presence of the volatile taint compounds, an important finding was that the electronic nose indicated significant differences between test and control samples when panellists performing sensory evaluation did not. This also correlated to an interval when enumeration on OSA illustrated no A. acidoterrestris after several days of inoculation and incubation and BAM and KM did. Without implying that the electronic nose has no drawbacks, it has proved, in this instance to be a simple and easy piece of equipment to use. It can be used to detect taints produced under simulated spoilage conditions at reduced analysis times, levels of expertise, cost and energy.