Prevalence, concentration, spoilage, and mitigation of Alicyclobacillus spp. in tropical and subtropical fruit juice concentrates

The presence of Alicyclobacillus in fruit juices and concentrates poses a serious problem for the juice industry. This study was undertaken to determine the (i) prevalence, concentration, and species of Alicyclobacillus in tropical and subtropical concentrates; (ii) efficacy of aqueous chlorine dioxide in reducing Alicyclobacillus spp. spores on tropical and subtropical fruit surfaces; and (iii) fate of and off-flavor production by Alicyclobacillus acidoterrestris in mango and pineapple juices. One hundred and eighty tropical and subtropical juice concentrates were screened for the presence and concentration of Alicyclobacillus spp. If found, the species of Alicyclobacillus was determined by 16S rDNA sequencing and analysis with NCI BLAST. Of these samples, 6.1% were positive for Alicyclobacillus, and nine A. acidoterrestris strains and two Alicyclobacillus acidocaldarius strains were identified. A five-strain cocktail of Alicyclobacillus spp. was inoculated onto the surface of fruits (grapefruit, guava, limes, mangoes, oranges and pineapple), which were then washed with 0, 50, or 100 ppm aqueous chlorine dioxide. Significant reductions due to chlorine dioxide were only seen on citrus fruits. A five-strain cocktail of A. acidoterrestris was inoculated into mango and pineapple juices. Microbial populations were enumerated over a 16-day period. Aroma compounds in the juice were analyzed by GC-olfactometry (GC-O) and confirmed using GC-MS. GC-O of mango juice identified previously reported medicinal/antiseptic compounds. GC-O of pineapple juice revealed an unexpected “cheese” off-aroma associated with 2-methylbutyric acid and 3-methylbutyric acid.     

Isolation,identification and typification of Alicyclobacillus acidoterrestris and Alicyclobacillus acidocaldarius strains from orchard soil and the fruit processing environment in South Africa

Alicyclobacillus acidoterrestris and Alicyclobacillus acidocaldarius are thermo-acidophilic, non-pathogenic, spore-forming bacteria that can survive the typical heat processing of fruit juices and concentrates. Bacterial endospores then germinate, grow and cause spoilage of acid food products. Species of Alicyclobacillus were isolated from orchard soil and a fruit concentrate production factory in South Africa. Preliminary identification of the isolates was based on morphological, biochemical and physiological properties. Identification at species level was done by PCR amplification using genus-specific primers and 16S ribosomal RNA (rRNA) gene sequencing. The majority of isolates belonged to the species A. acidoterrestris, but A. acidocaldarius was also isolated and identified. As far as we could determine, this is the first report of the isolation of A. acidoterrestris from wash water and soil outside a fruit processing plant, as well as the isolation of A. acidocaldarius from vinigar flies. The genotypic relatedness between strains of A. acidoterrestris and between strains of A. acidocaldarius was determined by RAPD-PCR. Sixteen isolates identified as A. acidoterrestris grouped into four clusters based on RAPD-PCR banding patterns, suggesting that they belong to at least four genotypic groups. Three isolateT:/PGN/ELSEVIER/YFMIC/web/00001155/s identified as A. acidocaldarius gave three unique banding patterns.     

Alicyclobacillus acidoterrestris in pasteurized exotic Brazilian fruit juices: Isolation,genotypic characterization and heat resistance

In this study, the population of Alicyclobacillus spp. was estimated in pasteurized exotic Brazilian fruit juices using the most probable number (MPN) technique followed by biochemical tests. Pasteurized passion fruit (n = 57) and pineapple (n = 50) juices were taken directly from Brazilian manufacturers. While Alicyclobacillus spp. was isolated from passion fruit juice, the microorganism was not found in any pineapple juice samples. A higher incidence of Alicyclobacillus was observed in samples taken in June and July (dry months in Brazil) in comparison to the other months (March, April, May and August), and the highest Alicyclobacillus counts were recovered from these samples(>23 MNP/100 mL). Sixteen (n = 16) Alicyclobacillus strains were typed using the randomly amplified polymorphic DNA method (RAPD-PCR). RAPD-PCR revealed great genetic similarity between the passion fruit juice strains and Alicyclobacillus acidoterrestris DSM 2498. The heat resistance of three isolates was determined, and the mean D_95°  (1.7 min) and z (7.6 °C) values in the passion fruit juice were not significantly different (p > 0.05) from those obtained for the DSM 2498 strain (D_95°  = 1.5 min and z = 7.1 °C). This is the first report on the isolation of A. acidoterrestris from exotic fruit juices such as passion fruit juice. It is worth pointing out the importance of applying good agricultural practices in the field and applying controls for the fruit selection and washing steps, as well as controlling the time/temperature conditions for pasteurization so as to reduce the incidence and chances of A. acidoterrestris spoilage in these juices.     

Effect of single and combined UV-C and ultra-high pressure homogenisation treatments on inactivation of Alicyclobacillus acidoterrestris spores in apple juice

Alicyclobacillus acidoterrestris is a spore-forming bacterium that can survive thermal pasteurization and acidic conditions. It produces changes in the odour and flavour of fruit juices leading to economical loses. A. acidoterrestris CECT 7094 spores were inoculated in clarified and cloudy apple juices (Golden delicious var.) in the range of 5–6 log₁₀ spores/mL and submitted to different short-wave ultraviolet light (UV-C) doses (7.2–28.7 J/mL) and ultra-high pressure homogenisation (UHPH) treatments (100–300 MPa), including their combination. A. acidoterrestris could be inactivated in clarified apple juice at a level of 4.8 log₁₀ CFU/mL by a 300 MPa-UHPH treatment when the inlet temperature was 80 °C. UV-C treatments showed to be more efficient achieving a lethality of 5.5 log₁₀ CFU/mL with a dose of 21.5 J/mL at 20 °C. In cloudy apple juice (2357 NTU) UV-C treatments were less efficient with a maximum lethality of 4.07 CFU/mL after a dose of 28.7 J/mL. A previous application of UHPH contributed with UV-C to obtain higher reductions of A. acidoterrestris spores at the doses of 14.3 and 21.5 J/mL compared with UV-C single treatments. On the other hand, this previous treatment also changed the properties of particles in the matrix which apparently reduced the effectiveness of UV-C at 28.7 J/mL.     

Use of a multivariate approach to assess the incidence of Alicyclobacillus spp. in concentrate fruit juices marketed in Argentina: results of a 14-year survey

The purpose of this study was to determine the incidence of Alicyclobacillus spp. in fruit/vegetable juices (concentrated pulps and clarified and non-clarified juices) marketed in Argentina between 1996 and 2009.The presence of Alicyclobacillus was determined in a total of 8556 samples of fruit and vegetable juices (apple, pear, grape, peach, blend of juices, tangerine, pineapple, orange, mango, plum, guava, apricot, lemon, banana, kiwi, carrot, strawberry, grapefruit, and beetroot) collected in seven Argentinean provinces. Multiple factor analysis (MFA) was carried out on a data matrix that contained the percentage of positive samples, type of juice, raw material and production year.Except for kiwi and orange, Alicyclobacillus was found in juices from all the evaluated raw materials. The highest percentage of positive samples was found for beetroot, strawberry, banana, peach, mango, carrot and plum juices. The percentage of positive samples for these juices ranged from 100% to 24%.Furthermore, the application of multivariate techniques provided an insight on the relationship between the incidence of Alicyclobacillus and production variables. This approach enabled the identification of the most relevant variables that increased the percentage of positive samples among the juices, which could help in developing strategies to avoid the incidence of this bacterium.By means of hierarchical cluster analysis seven groups (clusters) of juices which showed different percentages of positive samples for Alicyclobacillus spp. were identified. This analysis showed that pineapple, peach, strawberry, mango and beetroot juices had higher rates of positivity for Alicyclobacillus than the rest of the evaluated juices. MFA analysis also showed that some clear relationships could be highlighted between the percentage of samples positive for Alicyclobacillus and five types of fruit juices (strawberry, beetroot, grapefruit, pineapple and mango). It was observed that a large proportion of juices produced in 2000, 2005 and 2008 were located in clusters with higher incidence of Alicyclobacillus spp., whereas a larger proportion of clarified concentrate juice and concentrate pulp samples showed higher probability of incidence of Alicyclobacillus in these products. Data presented in this study brings a contribution to the ecology of Alicyclobacillus in fruit/vegetable juices marketed in Argentina. This information would be useful to enhance the microbiological stability of fruit juices regarding the presence of Alicyclobacillus spp.     

Isolation of Alicyclobacillus and the influence of different growth parameters

Alicyclobacillus species are thermo-acidophilic, endospore-forming bacteria that are able to survive pasteurisation and have been implicated in a number of spoilage incidents involving acidic foods and beverages. The aim of this study was to compare three isolation methods used for the detection of Alicyclobacillus acidoterrestris and to investigate the influence of incubation temperature on the growth of A. acidoterrestris and A. acidocaldarius. Peach juice samples inoculated with A. acidoterrestris K47 were analysed using either the International Federation of Fruit Juice Producers (IFU) Method No. 12 (Method A), which involved spread plating onto Bacillus acidoterrestris (BAT) agar at pH 4.0; Method B, which involved pour plating using potato dextrose agar (PDA) at pH 3.7; or Method C, which made use of membrane filtration followed by incubation on K agar at pH 3.7. The performance of the three methods differed significantly, with the IFU Method No. 12 recovering the highest percentage of cells at 75.97%, followed by Method B at 66.79% and Method C at 3.43%. These findings strengthen the proposal of the IFU for the use of the IFU Method No. 12 as a standard international method for the detection of Alicyclobacillus. To investigate the effect on growth of different incubation temperatures A. acidoterrestris (three strains) and A. acidocaldarius (two strains) were incubated at either 45 °C or 25 °C. Growth at 25 °C was slower and maximum cell concentrations were lower (1 × 10⁵-10⁶ cfu/mL compared to 1 × 10⁷-10⁸ cfu/mL) than at 45 °C for A. acidoterrestris. A. acidocaldarius was unable to grow at 25 °C and cell concentrations decreased by 1-2 logs. Since a growth temperature of 25 °C could not inhibit growth of A. acidoterrestris, cooling to room temperature (20°-25 °C) is not an effective control measure for A. acidoterrestris inhibition.     

Discrimination of <Emphasis Type="Italic">Alicyclobacillus</Emphasis> Strains by Lipase and Esterase Fingerprints

Alicyclobacillus spp. have recently become a major issue in the fruit juice industry due to its implication in the spoilage of pasteurized juices/beverages. In this study, lipase and esterase fingerprints of 37 Alicyclobacillus strains and 14 Bacillus strains in apple juice were collected using a five-substrate cocktail including four C8-esters and one C4-diester. Characterization of each strain could be finished in a single assay followed by high-performance liquid chromatography (HPLC) analysis for product separation and quantification. The obtained lipase and esterase fingerprints were highly species-dependent. Differences between the relative peak areas of each product for different species were significant. Coupled to chemometric techniques including principal component analysis (PCA) and stepwise linear discriminant analysis (SLDA), the lipase and esterase profiles led to 100 % of correct species identification for Alicyclobacillus and Bacillus strains. This approach is promising, reliable, and with good repeatability and could be used as an alternative tool to discriminate Alicyclobacillus spp. in fruit juices.     

Growth inhibition and inactivation of Alicyclobacillus acidoterrestris endospores in apple juice by hyperbaric storage at ambient temperature

Control of endospores of Alicyclobacillus acidoterrestris in pasteurized apple juice using hyperbaric storage at 18 to 23 °C was compared to storage at atmospheric pressure and 18 to 23 °C, as well as refrigeration at ~4 °C for up to 30 days. The juice samples were inoculated with approximately 1 × 10⁵ CFU/mL spores. The juice spoiled quickly at atmospheric pressure and ambient temperature, while under refrigeration spore levels remained unchanged for 30 days. Hyperbaric storage of inoculated apple juice at 25, 50 and 100 MPa at 18 to 23 °C resulted in spore inactivation at more rapid rates as pressure magnitudes increased, reaching levels below the detection limit of 10 CFU/mL at 50 and 100 MPa. In highly acid foods such as apple juice, hyperbaric storage at pressures ≤100 MPa and ambient temperature was effective in inactivating spores of A. acidoterrestris for periods up to 30 days.These results indicate hyperbaric storage at ambient temperature as a clearly more efficient preservation procedure to control the development of A. acidoterrestris endospores, compared to ambient temperature and refrigerated storage, in highly acidic foods as apple juice.     

Phylogenetic and spectroscopic analysis of Alicyclobacillus isolates by 16S rDNA sequencing and mid-infrared spectroscopy

Alicyclobacillus spp. are a group of thermophilic, acidophilic, spore-forming bacteria, some of which cause spoilage in commercially pasteurized fruit and vegetable juice products including apple, orange, tomato and carrot juice. In this study, we characterized seven isolates of Alicyclobacillus (14-2, KF, A-Gala 2-1, C-Fuji 6, Gala 9-2, 1016, 18-1) and a reference strain A. acidiphilus DSM 14558 by a 16S rDNA sequencing technique and Fourier transform infrared (FT-IR) spectral features. The degree of similarity between Alicyclobacillus isolates based upon phylogenetic analysis of sequence data and multivariate statistical analysis of FT-IR spectral data was determined. Results from both methods were consistent, suggesting that a combination of methods targeting both 16S rDNA and mid-infrared spectroscopic signatures may be a suitable and more accurate approach to either identify or discriminate Alicyclobacillus isolates from fruit and vegetable juice products.     

Development of Polyclonal Antibody‐Based Indirect Enzyme‐Linked Immunosorbent Assay for the Detection of Alicyclobacillus Strains in Apple Juice

Abstract: A sort of specific polyclonal anti‐Alicyclobacillus antibody was generated by immunizing New Zealand white rabbits, and a sensitive indirect enzyme‐linked immunosorbent assay (ELISA) was developed for Alicyclobacillus detection in apple juice. A set of experimental parameters such as concentration of antigen, dilutions of the antibody and goat anti‐rabbit IgG‐horseradish peroxidase conjugate, selection of the blocking reagent, incubation time, and temperature was optimized. The cross‐reactivity of the antibody was evaluated by ELISA and the result was consistent with Western blot analysis. The detection limit of the ELISA was about 10⁵ colony forming units (CFU)/mL in apple juice samples. Samples were detected by ELISA and conventional culture method, and the ELISA results gave a good agreement with the results obtained by plating on Alicyclobacillus acidoterrestris medium agar. ELISA takes a total detection time of 6 to 7 h, which is less than the time of conventional techniques requiring more than 24 to 48 h. These results indicated that the established ELISA was a potential useful analytical method for detection of Alicyclobacillus in apple juice.     

Alicyclobacillus acidoterrestris spores in fruit products and design of pasteurization processes

Alicyclobacillus acidoterrestris is a thermoacidophilic, non-pathogenic and sporeforming bacterium which has been found in commercial pasteurized fruit juices in the past. Only few and recent studies were available in the literature, since only in 1984 Cerny et al. [Cerny, G., Hennlich, W., & Poralla, K. (1984). Fruchtsaftverderb durch bacillen: isolierung und charakterisierung des verderbserregers. Z. Lebensmitt. Unters. Forsch. 179, 224–227] reported a spoiled aseptically packaged apple juice with A. acidoterrestris and in 1987 Deinhard et al. [Deinhard, G., Blanz, P., Poralla, K., & Altan, E. (1987). Bacillus acidoterrestris sp. nov., a new thermotolerant acidophile isolated from different soils. Systematic and Applied Microbiology, 10, 47–53] named first this species. Detection and identification methods for A. acidoterrestris were reviewed and data regarding heat resistance of spores and growth in fruits were collected. Finally, a new methodology to design pasteurization processes for high acidic fruit products is presented.     

Isolation and identification of Alicyclobacillus acidocaldarius by 16S rDNA from mango juice and concentrate

In this study we investigate the spoilage of ultra high temperature UHT mango juice as well as a carbonated fruit juice blend to identify organisms contributing to the spoilage. The mango concentrate, the final product, as well as the other ingredients used during manufacturing, were tested for the presence of Alicyclobacillus by polymerase chain reaction (PCR) and sequencing analyses. Microbiological examination of the mango pureé and spoiled fruit juices, using YSG agar [yeast extract 2 g, glucose 1 g, soluble starch 2 g, pH 3.7 (adjust with 2N H₂SO₄), H₂O 1000 mL, bacto agar 15 g] incubated at 55 °C, detected sporeforming, acid dependent and thermotolerant bacteria. The hyper variable region of the 16S rDNA was amplified. The nucleotide sequence of the PCR fragments was determined using the ABI Prism 310 automated DNA sequencer and the collected sequencing data were analysed and compared with the non‐redundant database using NCBI‐BLAST. Alicyclobacillus acidocaldarius were isolated and identified by 16S rDNA gene sequences analyses. The results indicated that the mango purèe, as well as the final product of mango juice and the fruit juice blend, were positive for Alicyclobacillus. The preventative measures of low pH, pasteurization of mango juice and the subsequent use of aseptic packaging were not regarded as sufficient to prevent the outgrowth of Alicyclobacillus spoilage organisms.     

Spoilage by Alicyclobacillus Bacteria in Juice and Beverage Products: Chemical,Physical, and Combined Control Methods

Alicyclobacillus is a genus of spoilage bacteria causing contamination of juices and other beverage products that cannot easily be contaminated by other microbes because of their high acid contents. During the last 4 decades since the first species of Alicyclobacillus was isolated in 1967, Alicyclobacillus has become a major concern to the global juice and beverage industries, and many promising methods have been developed and applied to control them. After introducing the history and general characteristics of Alicyclobacillus, as well as their heat resistance and spoilage, this review focuses on the control methods against Alicyclobacillus, including chemical and physical methods and combined methods. All these control methods show inhibitory or killing effects against Alicyclobacillus to some extent and, moreover, some of them have been put to use in the juice and beverage industries for decades and shown to be quite effective, although further developments can be achieved and new methods are constantly being established and investigated. Although it is difficult to compare the effects with one another among these methods because of the different experimental conditions in different reports, some of them, such as the treatments of nisin and high hydrostatic pressure, are well studied and proved to have extensive application prospect. The inhibitory factors, test strains and media, and especially the detailed experimental conditions and main results of these control methods are summarized here. The limitations of some methods mainly relating to the changes of products’ sensory qualities are also presented.     

GC–MS quantification and sensory thresholds of guaiacol in orange juice and its correlation with Alicyclobacillus spp.

Guaiacol is a profoundly negative taint/off-flavour produced by the increasingly common microbial contamination of fruit juices with Alicyclobacillus spp. The objectives of this study included: determining sensory thresholds for guaiacol in orange juice (OJ), developing an analytical method whose detection limits were equivalent to sensory thresholds and determining levels of Alicyclobacillus spp. that would produce detectable levels of guaiacol. A 12 member trained panel was used to establish guaiacol detection and recognition thresholds. Guaiacol ortho and retro nasal detection thresholds in OJ were 0.70 and 0.53 μg/l respectively. Odour recognition threshold was 2.0 μg/l. A SPME GC–MS Selected Ion Monitoring (SIM) procedure was optimised to achieve analytical detection limits of 0.5 μg/l. Optimum guaiacol detection limit was achieved using only responses from m/z 109 and 124. Ion ratios (m/z 109/124) and linear retention index value matching were used to confirm the identification of guaiacol. Quantification was achieved using peak areas from standard guaiacol additions in orange juice between 0.5 and 100 μg/l. Alicyclobacillus growth of 2.2 log CFU/ml in OJ produced just detectable levels (0.7 μg/l) of guaiacol.     

Antibacterial activity and mechanism of cinnamic acid and chlorogenic acid against Alicyclobacillus acidoterrestris vegetative cells in apple juice

The aim of this study was to investigate the antibacterial activity and mechanism of cinnamic acid and chlorogenic acid against Alicyclobacillus acidoterrestris. Minimum inhibitory concentrations of cinnamic acid and chlorogenic acid were 0.375 and 2.0 mg mL⁻¹, and the minimum bactericidal concentrations were 0.50 and 4.0 mg mL⁻¹, respectively. The apple juice ingredients had little influence on the inactivation of A. acidoterrestris. After treatment with cinnamic acid and chlorogenic acid, the morphology of A. acidoterrestris cells were severely destroyed; the leakage of nucleic acids and proteins increased significantly. SDS-PAGE investigation of bacterial proteins proved that the loss of soluble proteins was obvious as well. These results demonstrated that cinnamic acid and chlorogenic acid exerted their antibacterial activity mainly by an action mode of membrane disruption. This study provides an alternative method for the control of A. acidoterrestris-related spoilage in the fruit juice/beverage industry.     

Discrimination between Bacillus and Alicyclobacillus Isolates in Apple Juice by Fourier Transform Infrared Spectroscopy and Multivariate Analysis

Alicyclobacillus is a causative agent of spoilage in pasteurized and heat‐treated apple juice products. Differentiating between this genus and the closely related Bacillus is crucially important. In this study, Fourier transform infrared spectroscopy (FT‐IR) was used to identify and discriminate between 4 Alicyclobacillus strains and 4 Bacillus isolates inoculated individually into apple juice. Loading plots over the range of 1350 and 1700 cm^?1 reflected the most distinctive biochemical features of Bacillus and Alicyclobacillus. Multivariate statistical methods (for example, principal component analysis and soft independent modeling of class analogy) were used to analyze the spectral data. Distinctive separation of spectral samples was observed. This study demonstrates that FT‐IR spectroscopy in combination with multivariate analysis could serve as a rapid and effective tool for fruit juice industry to differentiate between Bacillus and Alicyclobacillus and to distinguish between species belonging to these 2 genera.     

Moulds and yeasts in fruit salads and fruit juices

Thirty-eight fruit salad samples including cantaloupe, citrus fruits, honeydew, pineapple, cut strawberries and mixed fruit salads, and 65 pasteurized fruit juice samples (apple, carrot, grapefruit, grape and orange juices, apple cider, and soy milk) were purchased from local supermarkets in the Washington, DC area and tested for fungal contamination. The majority of fruit salad samples (97%) were contaminated with yeasts at levels ranging from <2.0 to 9.72 log10 of colony forming units per gram (cfu/g). Frequently encountered yeasts were Pichia spp., Candida pulcherrima, C. lambica, C. sake, Rhodotorula spp., and Debaryomyces polymorphus. Low numbers of Penicillium spp. were found in pineapple salads, whereas Cladosporium spp. were present in mixed fruit and cut strawberry salads. Twenty-two per cent of the fruit juice samples tested showed fungal contamination. Yeasts were the predominant contaminants ranging from <1.0 to 6.83 log10 cfu/ml. Yeasts commonly found in fruit juices were C. lambica, C. sake, and Rhodotorula rubra. Geotrichum spp. and low numbers of Penicillium and Fusarium spp. (1.70 and 1.60 log10 cfu/ml, respectively) were present in grapefruit juice.     

Inactivation of Spoiling Yeasts of Fruit Juices by Pulsed Ultrasound

This paper reports on the use of ultrasound (US) as a suitable strategy to control the growth of spoiling yeasts in fruit juices. In a first phase, US technique was tested towards Saccharomyces cerevisiae inoculated in different juices (strawberry, orange, apple, pineapple and red-fruits): the treatment was performed by modulating the level of the power (20–60 %), the duration of the treatment (2–6 min) and the pulse (2–6 s), according to a fractional design. Then, the best treatment was applied against some other spoiling yeasts (Pichia membranifaciens, Wickerhamomyces anomalus, Zygosaccharomyces bailii, Zygosaccharomyces rouxii, Candida norvegica). Finally, a challenge test for a commercial beverage (red-fruit juice), inoculated with Z. bailii and containing a citrus extract, was conducted evaluating the effect of US on the sensory scores of the beverage. The results showed that the effect of US was mainly influenced by the power and the duration of the treatment; on the other hand, the effect of pulse was less significant and slight. The highest reduction of S. cerevisiae was found in the following combination of the design: power 60 %/time 4 min/pulse 2 s and power 60 %/time 6 min/pulse 6 s; this result was confirmed for the other spoiling yeasts. US and citrus extract could be combined to prolong the shelf life of the red-fruit juice and control the growth of Z. bailii. The two hurdles exerted a different role and acted in different times: US reduced the initial contamination, whilst citrus extract controlled the yeast within the storage.     

Effect of Ultrafiltration Combined with High-Pressure Processing on Safety and Quality Features of Fresh Apple Juice

Ultrafiltration (UF, 0.05 μm) with a ceramic membrane was combined with high-pressure processing (HPP) at 500 MPa/6 min and high-temperature short time (HTST) at 110 °C/8.6 s to process fresh apple juice. The aim of this study was to compare the effect of UF + HPP and UF + HTST on quality features of fresh apple juice and analyze the quality changes of the juice treated by UF + HPP and stored during 60 days at 4 °C. Applying UF, total plate count (TPC) and yeasts and molds (Y&M) significantly decreased by 0.29 and 0.28 log cycle, total phenols and ascorbic acid decreased by 33.50 and 26.52 %, and antioxidant capacity, using the DPPH and FRAP assay, significantly decreased by 26.40 and 25.37 %. Meanwhile, the juice clarity was 99.75?±?0.07 % and seven aroma compounds were changed. TPC and Y&M in juices treated by UF + HPP and UF + HTST were <1 log cycle. When compared to the juice treated by UF + HTST, the juice treated by UF + HPP showed lower browning degree and higher total phenols and clarity, and retained seven main volatile aroma compounds. Fresh apple juice processed by UF + HPP was microbiologically safe (TPC <1.8 log cycles and Y&M <1 log cycle) during 60 days of storage at 4 °C. The first-order model was a suitable model for all quality parameters of refrigerated fresh apple juice; however, rate constant k of first-order model was between ?0.0157 and 0.0350, showing the quality features of the refrigerated juice was stable.     

Occurrence of Alicyclobacillus in the fruit processing environment--a review

Concentrated fruit products have a significant place in modern consumption markets and are valuable semi-prepared food components to the bakery, dairy, confectionary, canning, baby food, frozen food, distilling and beverage industries. There is continuous pressure on the beverage industry to improve the quality of concentrated fruit products in order for reconstituted fruit beverages to compete with beverages that are made from fresh fruits. In recent years, Alicyclobacillus spp. have become a major concern to the beverage industry worldwide as many high-acid, concentrated fruit products have been found to be contaminated with these spoilage microbes. The thermo-acidophilic nature of alicyclobacilli and highly resistant endospores allows for their survival during the production of concentrated fruit products. Under favourable conditions, endospores can germinate and multiply to numbers high enough to cause spoilage and product deterioration through the production of chemical taint compounds. It is imperative to understand the nature of Alicyclobacillus within the fruit concentrate processing environment so as to develop effective control strategies and to prevent spoilage in juice and beverage products that are reconstituted from fruit concentrates. This paper reviews the occurrence of alicyclobacilli in the fruit processing environment, control measures, as well as detection, identification and standardised test methods that are currently used for Alicyclobacillus in concentrated fruit products.