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.     

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.     

Identification and haplotype distribution of Alicyclobacillus spp. from different juices and beverages

Alicyclobacillus spp. is an important thermoacidophilic, spore-forming spoilage bacterium that is a major concern for beverage and juice industries. In order to develop effective control strategies and adequately address the prevalence of contamination sources, it is necessary to characterize Alicyclobacillus' ecology in fruit, juice and beverage production and processing environments.Alicyclobacillus spp. isolates were collected from juice, beverage, ingredients, and environmental samples over a period of ten years. A total of 141 isolates were characterized as Alicyclobacillus spp. by 16S rRNA analysis and the most frequently isolated species was found to be Alicyclobacillus acidoterrestris (45%), A. acidocaldarius subsp. acidocaldarius (30%), and A. acidocaldarius (11%).The majority of thermotolerant sporeformers isolated from apple juices and concentrates was found to be A. acidoterrestris (24 out of 36 total apple isolates); while A. acidoterrestris was most frequently associated with citrus, citrus concentrates, and their associated environments, isolated by University of Florida (UF) (15 out of total 28 UF citrus isolates). However, A. acidocaldarius and subsp. acidocaldarius were frequently isolated by Cornell University (CU) (29 out of 35 CU citrus isolates), from citrus juices made from concentrate. Four major haplotypes of Alicyclobacillus spp. were identified based on the 16S rRNA gene sequencing from the 141 isolates tested. The Allelic Types (ATs) matched the phylogenetic analysis grouping of the different Alicyclobacillus spp. based on the isolation source.Our results suggest a predisposition for certain ATs of Alicyclobacillus spp. depending on juice or ingredient isolation source.     

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.     

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.     

Heat resistance of Neosartorya fischeri in various juices

Heat resistance of Neosartorya fischeri was studied in three different juices (apple, pineapple and papaya). The optimum heat activation temperature and time for the ascospores of the N. fischeri (growth for 30 days at 30 °C) was 85 °C for 10 min. Of the three juices tested, apple juice exhibited maximum 1/k values at 80, 85 and 90 °C (208.3, 30.1 and 2.0 min, respectively). The 1/k values for papaya juice (129.9, 19.0 and 1.9 min) and pineapple juice (73.5, 13.2 and 1.5 min) decreased with acidity and °Brix/acidity (ratio) level. The Z^* values for apple, papaya and pineapple juices were 5, 5.5 and 5.9 °C, respectively. The sterilization F values (4-log reduction) for apple, pineapple and papaya juices were 56.3, 38.0 and 7.2 s, respectively. Considering the thermal treatments commercially applied to pineapple (96 °C/30 s) and apple juices (95 °C/30 s), it is concluded that such treatments will not guarantee that less than 1 ascospore in each set of 10³ packs survive. Only the treatment applied to papaya juice (100 °C/30 s) will be sufficient because the F value is less than 30 s.     

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.     

UV-C treatment of juices to inactivate microorganisms using Dean vortex technology

A laboratory-scale UV-C treatment device based on Dean vortex technology was tested for its potential to inactivate spoilage microorganisms in cloudy fruit juices. A log 5 and log 6 reduction could be achieved by inactivating Lactobacillus plantarum BFE 5092 and Escherichia coli DH5α in naturally cloudy apple juice at 1.9 and 7.7 kJ/L, respectively. A treatment with 9.6 kJ/L led to an approximately log 4 inactivation of Saccharomyces cerevisiae DSM 70478 and Alicyclobacillus acidoterrestris DSM 2498. The effects of possible influencing parameters such as optical density, turbidity and viscosity were analyzed with regard to the efficiency of the UV-C treatment. The optical density based on dissolved compounds appeared to be the most important factor which influenced the bacterial inactivation efficiency. Cell counts of L. plantarum BFE 5092 could be reduced in quarter-strength Ringer’s solution adjusted with dye from an initial level of approximately 1 × 10⁸-1 × 10¹ cfu/mL at an optical density (254 nm) of 20 at 9.6 kJ/L. Only a log 1.5 reduction, however, could be achieved at an optical density (254 nm) of 140 using the same UV-C treatment. Furthermore, no noticeable effect on inactivation could be determined by varying the turbidity or the viscosity of the juices investigated. An increasing flow rate and the consequently higher Dean number clearly improved the efficacy of the UV-C treatment. Thus, the inactivation of L. plantarum BFE 5092 in blood orange juice could be enhanced by an approximately 2.5-log reduction by increasing the Dean number from 32 to 256 at 7.7 kJ/L. The UV-C treatment using Dean vortex technology was shown in this study to effectively inactivate microorganisms even in cloudy juices. The optical density value seemed to be the exclusive determining factor on the efficiency of the UV-C inactivation of microorganisms based on Dean vortex technology, while the effect of suspended solids was negligible as a result of the efficient mixing by Dean vortices.     

The Effect of RF Treatment Combined with Nisin Against <Emphasis Type="Italic">Alicyclobacillus</Emphasis> Spores in Kiwi Fruit Juice

In the present study, the efficiency of the treatment of radio frequency (RF) combined with nisin against the spores of three strains of Alicyclobacillus from two species was examined with the response surface methods (RSM). The results showed that the combination of RF and nisin was effective on killing Alicyclobacillus acidoterrestris and Alicyclobacillus contaminans spores in kiwi fruit juice. The killing effect increased with the increase of temperature, holding time, and nisin concentration, but the increment was not significant at high levels of temperature and nisin concentration. The killing effect between A. acidoterrestris and A. contaminans strains was different in kiwi fruit juice. The strain with the highest killing effect was DSM 17975^T, followed by DSM 3923^T and C-ZJB-12-33. The reduction of spore number of DSM 17975^T at the central design points of RSM was about 2.4. Under experimental conditions, nisin showed hardly any effect on killing DSM 17975^T spores when combined with RF.     

Application of a multi-pass high-pressure homogenization treatment for the pasteurization of fruit juices

This work evaluates both the effects of a multiple-pass high-pressure homogenization treatment on the microbial inactivation of selected microbial strains (Saccharomyces cerevisiae, Lactobacillus delbrueckii, Escherichia coli) inoculated into commercial fruit juices (orange, red orange, pineapple) as well as the application of this non-thermal technology to the pasteurization of fresh juices (Annurca apple juice). The pressure level ranged from 50 to 250 MPa, the number of passes from 1 to 5 and the inlet temperature from 2 to 20 °C.Preliminary tests in distilled water showed that the efficiency of the multiple-pass treatment significantly depends on both the homogenizing pressure as well as the microbial species. The subsequent extension of the multiple-pass treatment to the inactivation of S. cerevisiae inoculated into three different fruit juices (orange, red-orange and pineapple juice) highlighted that the inactivation induced by the high pressure treatment did not depend on the properties of the tested juices and was not statistically different from inactivation in water (p value < 0.05). These findings were supported by the comparison of two different mathematical models used to fit the inactivation kinetics, whose fitting parameters were not significantly different for water and the fruit juices for any pressure level applied.Three homogenization passes at 150 MPa and 25 °C, which resulted to be optimal for yeast inactivation in fruit juices, were effective for the stabilization of the endogenous microbial load of fresh Annurca apple juice. The treated apple juice showed a minimum shelf-life of 28 days under refrigerated conditions, during which the natural qualities of the fresh juice were completely preserved.     

Bacterial spore inactivation at 45-65 °C using high pressure processing: Study of Alicyclobacillus acidoterrestris in orange juice

High pressure processing (HPP) is a new non-thermal technology commercially used to pasteurize fruit juices and extend shelf-life, while preserving delicate aromas/flavours and bioactive constituents. Given the spoilage incidents and economic losses due to Alicyclobacillus acidoterrestris in the fruit juice industry, the use of high pressure (200 MPa – 600 MPa) in combination with mild temperature (45 °C–65 °C) for 1–15 min, to inactivate these spores in orange juice were investigated. As expected, the higher the temperature, pressure and time, the larger was the A. acidoterrestris inactivation. The survival curves were described by the first order Bigelow model. For 200 MPa, D_45 °C = 43.9 min, D_55 °C = 28.8 min, D_65 °C = 5.0 min and z-value = 21.3 °C. At 600 MPa, D_45 °C = 12.9 min, D_55 °C = 7.0 min, D_65 °C = 3.4 min and z-value = 34.4 °C. Spores were inactivated at 45 °C and 600 MPa, and at 65 °C only 200 MPa was needed to achieve reduction in spore numbers.     

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.     

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.     

Heat resistance of the spoilage yeastsCandida pelliculosaandKloeckera apisand pasteurization values for some tropical fruit juices and nectars

Spoilage yeasts, identified asCandida pelliculosaandKloeckera apis, were isolated from fermented pasteurized pineapple juice, guava and passion fruit nectars, and studied for their heat resistance in relation to the pasteurization process of the beverages. The decimal reduction times (D_T) obtained between 55 and 60^°C forK. apisvaried from 1.41 to 2.49 min, and those forC. pelliculosaobtained between 60 and 75^°C varied from 1.03 to 4.90 min depending upon the beverage and the tested temperatures.C. pelliculosaseemed to be more heat resistant thanK. apis. Thez-values or the increase in temperature needed to reduce theD_Tby 10, determined between 60 and 75^°C forC. pelliculosa, were 31.75±0.03^°C (r²=0.91) in pineapple juice, 27.70±0.04^°C (r²=0.90) in passion fruit nectar and 34.84±0.02^°C (r²=0.998) in guava nectar. ForK. apis, thez-values determined between 55 and 60^°C were 21.88±0.07^°C (r²=0.97) in pineapple juice, 22.73±0.04^°C (r²=0.99) in guava nectar and 29.07±0.01^°C (r²=0.999) in passion fruit nectar. The minimal pasteurizing valuesP_T^zwere determined forC. pelliculosaat 70^°C and at 75^°C, at a pasteurization efficiency (logN₀/N) of 9. In pineapple juice,P^z₇₀=27.91 min andP^z₇₅=13.5 min would have be applied to destroy almost any vegetative forms of the yeast. In passion fruit nectar,P^z₇₀=20.25 min andP^z₇₅=9.27 min, and in guava nectarP^z₇₀=16.83 min would have be applied.     

Effects of tangential microfiltration and pasteurisation on the rheological,microbiological, physico‐chemical and sensory characteristics of sugar cane juice

The aim of this study was to determine the physico‐chemical, microbiological, rheological and sensory characteristics of sugar cane juice with passion fruit pulp (5% w/w) added and which had been submitted to microfiltration and pasteurisation (90 °C/30 s; 95 °C/30 s) processes. The results showed that the juices obtained after the microfiltration and pasteurisation processes had low microbial counts. The microfiltered juice showed a reduction in the soluble solids, acidity and vitamin C content (P = 0.05), while the pasteurised juice showed no change in these parameters (P > 0.05) in relation to the natural sugar cane juice, with the exception of vitamin C, which was not detected after the thermal treatment. In relation to the rheological properties, Newtonian behaviour was observed for both microfiltered and pasteurised juices. The sensory tests on a hedonic scale showed good acceptance of both juices (microfiltered and pasteurised).     

The impact of high power ultrasound for controlling spoilage by Alicyclobacillus acidoterrestris: A population and a single spore assessment

Consumer demand for healthier, microbiologically safe and stable, higher quality and minimally processed foods has increased in the last decades, promoting the application of non-thermal process technologies. Ultrasound processing is gaining attention because of its potential for improving quality and safety while retaining product flavor, texture, color and nutrient composition. Recently, Alicyclobacillus acidoterrestris has been recognized by manufacturers and processors in the fruit industry as a new type of thermoacidophilic, endospore-forming spoilage bacterium for commercialized fruit juices that can withstand the high temperatures applied in pasteurization process and spoil them producing taint compounds. Based on the above, the present study was undertaken to investigate the separate and combined effect of ultrasound treatment operating at 26 kHz, 90 μm, 200 W for 5, 10, 15 min and heat stress (at 80 °C for 10 min) on the recovery of A. acidoterrestris spores in K broth adjusted to pH = 4.5 with 25% (w/v) citric acid at temperatures between 35 and 45 °C at population and individual spore level. At the population level, statistically significant differences in average lag time of A. acidoterrestris spores from different treatments at 35 and 45 °C have been found. After the applied ultrasound (at 100% Amplitude for 10 min) and heat shock (at 80 °C for 10 min), the average lag time of A. acidoterrestris spores at 35 °C (7.24 ± 0.34 h) and 45 °C (6.38 ± 0.18 h) has been shown to be longer compared to the control at 35 °C (5.68 ± 0.36 h) and at 45 °C (2.87 ± 0.19 h), while for this combination, the maximum specific growth rate was not significantly different from the control samples. Additionally, the findings of this study have shown that λ among individual spores was greatly variable when they were treated by ultrasound (at 100% Amplitude for 10 min) and/or thermal treatment (at 80 °C for 10 min). The application of ultrasound and thermal treatment resulted in a more pronounced effect on median lag phase duration (25.74 h) than the heat shock (11.63 h) and affected both the position and the spread of the λ distributions of A. acidoterrestris spores. This work would help to better assess and optimize the proposed combined treatments, since ultrasound and thermal treatments could work in a synergistic way on delaying the lag time of the tested bacterial spores.     

Modelling the effect of temperature and water activity on the growth rate and growth/no growth interface of Byssochlamys fulva and Byssochlamys nivea

The purpose of the present study was to apply a modelling approach to define the growth rate and growth/no growth interface of Byssochlamys fulva and Byssochlamys nivea on a synthetic medium as a function of temperature and water activity. Both fungal species were grown on malt extract agar at different temperatures (10, 15, 20, 25, 30, 35, 40 and 45 °C) and a_w levels (0.88, 0.90, 0.92, 0.94, 0.96 and 0.99) for a period of 30 days. Growth responses were evaluated over time in terms of colony diameter changes. Growth data were fitted to the primary model of Baranyi and the resulting growth rates were further modeled as a function of temperature and water activity using the cardinal model with inflection (CMI) ( Rosso et al., 1993). A logistic regression quadratic polynomial model was also employed to predict the probability of growth over storage time. Estimated parameters for minimum, maximum and optimum temperatures for growth were 9.1 °C, 46.4 °C and 32.1 °C for B. fulva and 10.5 °C, 43.2 °C and 32.1 °C for B. nivea. The respective values for a_w were 0.893, 0.993 and 0.985 for B. fulva and 0.892, 0.992 and 0.984 for B. nivea. No growth was observed at 0.88 a_w regardless of temperature for both species, whereas B. nivea ascospores could not grow at 10 and 45 °C irrespective of a_w. Regarding growth boundaries, the degree of agreement between predictions and observations was >98% concordant for both species. The erroneously predicted growth cases were 1.4–4.2% false positive and 2.1–3.5% false negative for B. nivea and B. fulva, respectively. The developed logistic model was validated with two literature data sets as well as with data from independent experiments carried out on fruit juices. Validation results showed that agreement with literature data for growth was 25 out of 36 (69.4%) cases, whereas validation on fruit juice data failed in only 6 cases (5 false positives and 1 false negative) out of 128 cases.     

Storage quality of pineapple juice non-thermally pasteurized and clarified by microfiltration

Microfiltration (MF) is classified as a non-thermal process for the fruit juice industry. It could provide a better preservation of the phytochemical property and flavor of the juice. This work aimed to study the stability of phytochemical properties including vitamin C, total phenolic content, antioxidant capacity (2-Diphenly-1-picrylhydrazyl: DPPH, free radical scavenging capacity and Oxygen Radical Absorbance Capacity: ORAC assays), microbial and chemical–physical (color, browning index, pH and total soluble solid) properties of MF-clarified pineapple juice during storage at various temperatures (i.e. 4, 27, and 37 °C). The juices were clarified by microfiltration using hollow fiber module. The results showed that most of the phytochemical properties and soluble components were retained in the juice after microfiltration. No microbial growth was detected after 6 months of storage. The storage time and temperature did not affect total soluble solids and pH (P > 0.05). The color (L^*) of clarified juice stored at 4 °C was lighter than the juices stored at higher temperature levels (P < 0.05). The phytochemical properties and total phenol content of the juice significantly decreased as storage time and temperature increased (P < 0.05). Vitamin C content was the attribute that affected storage time and temperature most as indicated by reaction rate constant and activated energy. Storage of non-thermally pasteurized and clarified pineapple juice at 4 °C was the most suitable since it allowed the best quality preservation.     

Optimization of Tropical Juice Composition for the Spanish Market

ABSTRACT: The optimal proportions of 3 concentrate juices, passion fruit, pineapple, and banana, mixed with white grape juice were investigated by preferences mapping techniques. The objective was to establish the minimum level of each concentrated tropical juice necessary to add to a white grape juice so that it would be perceived as a mixture by consumers. A nonverbal test, analyzed by proxscal, demonstrated that decreasing an amount of concentrate juice from 4% to 1% would have no statistically significant effect on flavor recognition by consumers. Internal preference map for flavor and color acceptability revealed that a commercial mix of pineapple with white grape juice mix (used as control) was the most accepted juice, closely followed by an experimental mix of passion fruit with white grape juice. Principal component analysis revealed the importance of overall taste, overall odor, acid fruit aroma, acid taste, and pulp in the acceptability of these 3 juices.     

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.