Abstract
It was used reverse-transcriptase polymerase chain reaction (RT-PCR) followed by capillary electrophoresis on a microchip to probe the viability of Alicyclobacillus acidoterrestris spores after inactivation with saponin and heat. The method was shown to be suitable for the purpose, and was faster and more sensitive than the traditional plating technique, the standard of the food industry. The limits of quantification and detection were 0.0107 and 0.0039 ng µL-1 of amplified DNA, respectively. The correlation coefficient (r) between traditional plating and our method was 0.9977, which indicates an excellent correspondence between them. Consequently, it was possible to assess, with confidence, the viability of bacteria using the RT-PCR reaction for detection. It was evaluated the potential of this molecular method over traditional microbiology in the inactivation of A. acidoterrestris by saponin as an effective agent, potentiated by heat.
Highlights
The physical and chemical characteristics of concentrated orange juice combined with the effects of industrial thermal treatments usually inhibit most pathogens in this beverage.[1]
The purpose of this work was to correlate the plating technique[24] and the new reverse-transcriptase polymerase chain reaction (RT-polymerase chain reaction (PCR)) method coupled with microchip detection in the inhibition of A. acidoterrestris spores in orange juice by saponin and thermal treatments
It was noticed that the relative standard deviation (RSD) were higher for analyses performed on different days
Summary
The physical and chemical characteristics of concentrated orange juice combined with the effects of industrial thermal treatments usually inhibit most pathogens in this beverage.[1] the spoilage of fruit juices by the thermoacidophilic spore-forming bacterium Alicyclobacillus acidoterrestris has been observed on a worldwide level, with increasing beverage spoilage seen during the last two decades.[2,3] Fresh, concentrated, or even pasteurized orange juices are susceptible to spoilage by spores of A. acidoterrestris, which can survive the. Endospores represent the most difficult life form to inactivate since they show a great resistance to heat inactivation, and to chemical treatments.[7,8] Many compounds have been tested against A. acidoterrestris spores, with bacteriocins as the most studied preservatives. Nisin is more effective on spores than on cells; it seems to act at the stage of pre-germinant swelling, and at low concentrations (50‐100 IU mL-1), its effect is sporostatic rather than sporicidal.[9,10] Other bacteriocins have been proposed, such as enterocin AS‐48, extracted from
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