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Simultaneous monitoring of organic acids and sugars in fresh and processed apple juice by Fourier transform infrared-attenuated total reflection spectroscopy.

A combination of Fourier transform infrared spectroscopy (FT-IR) and chemometrics was used as a screening tool for the determination of sugars and organic acids such as sucrose, glucose, fructose, sorbitol, citric acid, and malic acid in processed commercial and extracted fresh apple juices. Prepared samples of synthetic apple juice in different constituent concentration ranges were scanned by attenuated total reflectance (ATR) accessory and the spectral region in the range between 950 and 1500 cm(-1) was selected for calibration model development using partial least squares (PLS) regression and principal component regression (PCR). The calibration models were successfully validated by high-performance liquid chromatography (HPLC) measurements against several commercial juice varieties as well as juice extracted from different apple varieties to provide an overall R2 correlation of 0.998. The present study demonstrates that Fourier transform infrared spectroscopy could be used for rapid and nondestructive determination of multiple constituents in commercial and fresh apple juices. Results indicate this approach to be a rapid and cost-effective tool for routine monitoring of multiple constituents in a fruit juice production facility.

Beverages↗

[Musculoskeletal symptoms and working postures in pear and apple orchard workers].

Musculoskeletal symptoms in pear and apple orchard workers were studied in relation to working postures, particularly arm elevation and head extension. The same forty-six female workers were examined three times; in May while thinning pears, in June while bagging pears, and in July while bagging apples. Musculoskeletal symptoms in the neck and shoulders were predominant in thinning and bagging pears when compared with bagging apples. When thinning pears, there was a significantly higher prevalence of complaints of stiffness and pain in the neck and shoulders, pain in the upper and lower back, muscle tenderness in the shoulders and forearm, and decreased back muscle power. When bagging pears, there was a significant difference in complaints of stiffness and pain in the neck and shoulders, muscle tenderness in the shoulders, and pain in motion of the neck joint. Pear tasks require more arm elevation and head extension than apple tasks. The working postures of arm elevation and head extension were considered to be a dominant causative factor in shoulder-neck disorders of the workers examined. The low back pain from thinning pears may be related with continuous backward bending of the back associated with the thinning work.

Agriculture↗

Monitoring codling moth (Lepidoptera: Tortricidae) with passive interception traps in sex pheromone-treated apple orchards.

Male and female codling moth, Cydia pomonella (L.), were monitored with passive interception traps (PI-traps) in apple orchards treated with sex pheromone dispensers. The proportion of mated females recaptured by PI-traps was significantly higher than the proportion released after the release of both sexes into a codling moth-infested orchard. However, no significant difference occurred between the proportion of mated females recaptured and released when only females were released into uninfested orchards. Replicated nine-tree apple plots situated either on the edge or in the center ofpheromone-treated apple orchards were monitored with PI-traps during first moth flight in 1995 and during both flights in 1996. Moths caught on PI-traps were predominately males. The first male moths were captured 7-10 d before females during the first flight in both years. Initial capture of virgin and mated females on PI-traps coincided in 1995. Mated females were captured 14 d after the first virgin females in 1996. The mean proportion of females that were mated ranged from 32 to 55% during the first flight and 85 to 92% during the second flight. Moth catch and fruit injury were significantly higher in the edge versus the center plots. The numbers of total and female moths caught with PI-traps were significantly correlated with fruit injury for each generation. The percentage of female moths caught on PI-traps that were mated was 32% lower and the mean oocyte load of all females was 42% higher in a pheromone-treated apple orchard than in the untreated crabapple grove monitored during May and June 1997.

Animals↗

Population dynamics of apple maggot (Diptera: Tephritidae) in south central Pennsylvania.

The apple maggot, Rhagoletis pomonella (Walsh), was monitored with baited yellow panels and red spheres in commercial orchards, abandoned orchards, and unsprayed backyard apple trees in 1998 and 1999. Apple maggot adults were captured in all apple habitats, but the capture levels in the abandoned orchards and unsprayed backyard trees tended to be higher than in the commercial orchards. Peak capture occurred between mid-July and late August in both years. Emergence cages seeded with infested fruits were used to investigate bivoltinism, which was observed in both years.

Animals↗

Prediction of valid acidity in intact apples with Fourier transform near infrared spectroscopy.

To develop nondestructive acidity prediction for intact Fuji apples, the potential of Fourier transform near infrared (FT-NIR) method with fiber optics in interactance mode was investigated. Interactance in the 800 nm to 2619 nm region was measured for intact apples, harvested from early to late maturity stages. Spectral data were analyzed by two multivariate calibration techniques including partial least squares (PLS) and principal component regression (PCR) methods. A total of 120 Fuji apples were tested and 80 of them were used to form a calibration data set. The influences of different data preprocessing and spectra treatments were also quantified. Calibration models based on smoothing spectra were slightly worse than that based on derivative spectra, and the best result was obtained when the segment length was 5 nm and the gap size was 10 points. Depending on data preprocessing and PLS method, the best prediction model yielded correlation coefficient of determination (r2) of 0.759, low root mean square error of prediction (RMSEP) of 0.0677, low root mean square error of calibration (RMSEC) of 0.0562. The results indicated the feasibility of FT-NIR spectral analysis for predicting apple valid acidity in a nondestructive way.

Algorithms↗

[Inter-laboratory study for validation of a Japanese official analytical method for determination of patulin in apple juice].

To validate a modified version of AOAC official method of analysis 995.10 as an official standard in Japan for determination of patulin in apple juice, an inter-laboratory study was performed in 11 laboratories using a non-contaminated sample, 2 naturally contaminated samples and 2 spiked samples of apple juice. For naturally contaminated apple juices, the relative standard deviations for repeatability and reproducibility were 3.2, 7.1% and 10.0, 21.7%, respectively. HORRAT values were 0.4, 0.9. The average recovery of patulin from spiked sample was 83.7%. The limit of quantification was calculated as 10 microg/kg. From these results, the method was thought to be suitable as an official standard for determination of patulin in apple juice in Japan.

Chromatography, High Pressure Liquid↗

Genome-Wide Characterization of the Apple HD-Zip IV Gene Family and Functional Validation of MdHDZIV3 Under PEG-Induced Osmotic Stress.

The homeodomain-leucine zipper IV (HD-Zip IV) transcription factor subfamily plays essential roles in epidermal development, cuticle formation, lipid metabolism, and environmental adaptation in plants. Despite its biological importance, the HD-Zip IV family has not been systematically characterized in apple (Malus domestica). Here, we identified 17 apple HD-Zip IV genes and named them MdHDZIV1-MdHDZIV17 based on their locations on the chromosomes. The 17 genes showed a nonuniform distribution on eight chromosomes, while the occurrence of both tandem and segmental duplications indicated that family expansion involved more than one duplication mechanism. All MdHDZIV proteins contained the conserved HD, LZ, START, and SAD domains but lacked the MEKHLA domain, consistent with typical HD-Zip IV structural features. Phylogenetic analysis classified MdHDZIV proteins into five groups together with HD-Zip IV members from Arabidopsis thaliana and rice, indicating evolutionary conservation of this subfamily. Collinearity and Ka/Ks analyses revealed that duplicated MdHDZIV gene pairs were mainly subjected to purifying selection. Promoter scanning revealed diverse cis-regulatory motifs associated with hormonal signaling, environmental stress, light response, and epidermal regulation, including ABRE, ARE, W-box, MYC, G-box, and L1-box motifs. Integration of transcriptomic profiling with qRT-PCR validation revealed pronounced tissue-dependent differences in the expression of MdHDZIV genes in leaf, fruit skin, and branch bark. Under PEG6000-induced osmotic stress and NaCl-induced salt stress, 10 candidate MdHDZIV genes displayed gene-specific and stress type-specific expression patterns, with MdHDZIV3 showing strong induction under PEG6000 treatment. Functional validation in apple calli showed that MdHDZIV3 overexpression enhanced PEG tolerance, increased fresh weight, elevated SOD and POD activities, and reduced MDA accumulation under osmotic stress. These findings provide a genome-wide framework for understanding the apple HD-Zip IV gene family.

abiotic stress↗

Detection of guaiacol produced by Alicyclobacillus acidoterrestris in apple juice by sensory and chromatographic analyses, and comparison with spore and vegetative cell populations.

Spoilage of fruit juice by Alicyclobacillus acidoterrestris is characterized by a distinct medicinal or antiseptic off odor attributed to guaiacol, a metabolic by product of the bacterium. Detection of low populations of A. acidoterrestris that would precede sensory detection of guaiacol would enable juice processors to select appropriate processing and storage conditions that would minimize or eliminate spoilage. The objective of this study was to determine the recognition threshold of guaiacol in apple juice by sensory analysis and the population of A. acidoterrestris and incubation time at 21 and 37 degrees C necessary for chemical detection of guaiacol. Commercially sterilized apple juice (pH 3.54 +/- 0.04, 11.3 +/- 0.3 degrees Brix) was inoculated with a five-strain mixture of A. acidoterrestris spores (2.98 log10 CFU/ml) and stored at 21 or 37 degrees C for up to 61 days. Using an experienced sensory panel and the forced-choice ascending concentration method of limits, the best estimate threshold (BET) for recognition of guaiacol added to uninoculated apple juice was 2.23 ppb. Numbers of A. acidoterrestris spores and cells in inoculated juice remained constant during the 61-day storage period; however, the panel detected (P < or = 0.01) guaiacol in juice stored at 37 degrees C within 8 days. At three of four sampling times ranging from 13 to 61 days at which the sensory panel detected (P < or = 0.001) guaiacol, concentrations of 8.1 to 11.4 ppb were detected by chromatographic analysis. The panel detected (P < or = 0.1 to P < or = 0.01) guaiacol in five samples stored at 21 to 37 degrees C for 8 to 61 days in which the compound was not detected by chromatographic analyses. It appears that guaiacol content in apple juice inoculated with A. acidoterrestris is not always correlated with numbers of cells, and the limit of sensitivity of chromatographic quantitation of the compound is higher than the BET.

Bacillus↗

Reduction of Escherichia coli O157:H7 counts on whole fresh apples by treatment with sanitizers.

The objectives of this study were to determine if washing of whole apples with solutions of three different sanitizers (peroxyacetic acid, chlorine dioxide, or a chlorine-phosphate buffer solution) could reduce a contaminating nonpathogenic Escherichia coli O157:H7 population by 5 logs and at what sanitizer concentration and wash time such a reduction could be achieved. Sanitizers were tested at 1, 2, 4, 8, and 16 times the manufacturer's recommended concentration at wash times of 5, 10, and 15 min. Whole, sound Braeburn apples were inoculated with approximately 1 x 108 or 7 x 106 CFU per apple, stored for 24 h, then washed with sterile water (control) or with sanitizers for the prescribed time. Recovered bacteria were enumerated on trypticase soy agar. Washing with water alone reduced the recoverable population by almost 2 logs from the starting population; this can be attributed to physical removal of organisms from the apple surface. No sanitizer, when used at the recommended concentration, reduced the recovered E. coli population by 5 logs under the test conditions. The most effective sanitizer, peroxyacetic acid, achieved a 5-log reduction when used at 2.1 to 14 times its recommended concentration, depending on the length of the wash time. The chlorine-phosphate buffer solution reduced the population by 5 logs when used at 3 to 15 times its recommended concentration, depending on wash time. At no concentration or wash time tested did chlorine dioxide achieve the 5-log reduction.

Buffers↗

Heat inactivation of Escherichia coli O157:H7 in apple juice exposed to chlorine.

Exposure of Escherichia coli O157:H7 to chlorine before heat treatment results in increased production of heat shock proteins. Current heating regimens for pasteurizing apple cider do not account for chlorine exposure in the wash water. This research determined the effect of sublethal chlorine treatment on thermal inactivation of E. coli O157:H7. D58-values were calculated for stationary-phase cells exposed to 0.6 mg/liter of total available chlorine and unchlorinated cells in commercial shelf-stable apple juice (pH 3.6). D58-values for unchlorinated and chlorine-exposed cells in buffer were 5.45 and 1.65 min, respectively (P < 0.01). Death curves of chlorine-exposed and unchlorinated cells in apple juice were not completely linear. Unchlorinated cells heated in apple juice exhibit a 3-min delay before onset of linear inactivation. Chlorine treatment eliminated this shoulder, indicating an overall loss of thermotolerance. The linear portion of each curve represented a small fraction of the total population. D58-values calculated from these populations are 0.77 min for unexposed cells and 1.19 min for chlorine-exposed cells (P = 0.05). This indicates that a subpopulation of chorine-treated cells is possibly more resistant to heat because of chlorine treatment. The effect of chlorine treatment, however, is insignificant when compared with the effect of losing the shoulder. This is illustrated by the time required to kill the initial 90% of the cell population. This is observed to be 3.14 min for unchlorinated versus 0.3 min for chlorine-exposed cells (P < 0.001). These observations indicate that current heat treatments need not be adjusted for the effect of chlorine treatment.

Animals↗

Hot water immersion to eliminate Escherichia coli O157:H7 on the surface of whole apples: thermal effects and efficacy.

The effect of hot water immersion on both the reduction of Escherichia coli O157:H7 on the apple surface and internal temperatures of the apple was assessed in this study. Microbial reductions were measured experimentally, whereas internal temperatures were calculated through a mathematical analysis of experimental heat transfer data obtained from the apples. A method was developed to provide a purely surface-based inoculation of E. coli O157:H7. Rinsing produced no reduction, and treatments at 80 and 95 degrees C produced reductions of more than 5 logs in 15 s or less. The heat transfer analysis based on experimental data was used to calculate surface heat transfer coefficients and predict temperatures throughout the apple. The analysis indicated a low heat transfer rate. Although it reduces thermal degradation, a low heat transfer rate precludes thermal-based reduction of any internalized microorganisms.

Colony Count, Microbial↗

Reduction of patulin during apple juice clarification.

Patulin is a mycotoxin produced by a number of molds involved in fruit spoilage. This compound is carcinogenic and teratogenic. Various methods are currently used to reduce the levels of patulin in apple juice, namely, charcoal treatment, chemical preservation (sulfur dioxide), gamma irradiation, fermentation, and trimming of fungus-infected apples. Many of these processes are expensive and time-consuming. Therefore, there is a need to find a convenient and economical process to control patulin levels. This study was undertaken to evaluate the effectiveness of several clarification processes for the reduction of patulin. Clarification was carried out on a laboratory scale. Apple pulp was spiked with patulin, pressed, and clarified using four different processes, namely, fining with bentonite, enzyme (pectinase) treatment, paper filtration, and centrifugation. Patulin was recovered from the clarified juice by liquid-liquid extraction, and solid-phase chromatography was used for sample clean-up prior to analysis by high-performance liquid chromatography (HPLC). The minimum detectable limit using HPLC was 20 microg/liter. Pressing followed by centrifugation resulted in an average toxin reduction of 89%. Total toxin reduction using filtration, enzyme treatment, and fining were 70, 73, and 77%, respectively. Patulin reduction was due to the binding of the toxin to solid substrates that was verified by analyzing the clarified juice as well as the filter cake, pellet, and sediment. The combined concentrations correlated to the spiked concentration. These results reveal that clarification was successful in the reduction of patulin levels in apple juice. However, clarification resulted in high levels of patulin in the pressed pulp after filtration and centrifugation, and this could be harmful if they are used as animal feeds.

Beverages↗

Inactivation of Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes on apples, oranges, and tomatoes by lactic acid with hydrogen peroxide.

The objective of this study was to develop a practical and effective method for inactivating or substantially reducing Escherichia coli O157:H7, Salmonella Enteritidis, and Listeria monocytogenes on apples, oranges, and tomatoes. Apples, oranges, and tomatoes were spot-inoculated with five-strain mixtures of E. coli O157:H7, Salmonella Enteritidis, and L. monocytogenes near the stem end and were submerged in sterile deionized water containing 1.5% lactic acid plus 1.5% hydrogen peroxide for 15 min at 40 degrees C. Inoculated samples treated with sterile deionized water at the same temperature and for the same duration served as controls. The bacterial pathogens on fruits subjected to the chemical treatment were reduced by >5.0 log10 CFU per fruit, whereas washing in deionized water decreased the pathogens by only 1.5 to 2.0 log10 CFU per fruit. Furthermore, substantial populations of the pathogens survived in the control wash water, whereas no E. coli O157:H7, Salmonella Enteritidis, or L. monocytogenes cells were detected in the chemical treatment solution. The sensory and qualitative characteristics of apples treated with the chemical wash solution were not adversely affected by the treatment. It was found that the treatment developed in this study could effectively be used to kill E. coli O157:H7, Salmonella Enteritidis, and L. monocytogenes on apples, oranges, and tomatoes at the processing or packaging level.

Colony Count, Microbial↗

Inactivation of Listeria monocytogenes Scott A 49594 in apple juice supplemented with cinnamon.

Normal (pH 3.7) and adjusted (pH 5.0) pasteurized apple juice containing cinnamon (0, 0.1, 0.2, and 0.3%) was inoculated with Listeria monocytogenes Scott A 49594 at 10(4) CFU/ml and stored at 5 and 20 degrees C for 7 days. Counts on tryptic soy agar (TSA), modified Oxford (MOX) medium, and thin agar layer (TAL) were determined at 1 h and 1, 3, and 7 days. The TAL method (MOX medium overlaid with TSA) was used for the recovery of injured cells. In apple juice, both at normal and adjusted pH, with any doses of cinnamon, no L. monocytogenes (a 4.6-log CFU/ml reduction) was detected after 1 h of storage at both temperatures, and no growth occurred at any points of storage. Therefore, cinnamon by itself (regardless of pH) had a pronounced killing effect. A further enrichment step with brain heart infusion agar showed that L monocytogenes was completely inactivated in apple juice stored at 20 degrees C, except in pH 5.0 samples with 0.1% of cinnamon. The TAL method was as effective as TSA in recovering injured cells of L. monocytogenes. Cinnamon considerably inactivates L. monocytogenes in apple juice and thus enhances the safety of this product.

Beverages↗

Polyphenol oxidase activity as a potential intrinsic index of adequate thermal pasteurization of apple cider.

In response to increasing concerns about microbial safety of apple cider, the U.S. Food and Drug Administration has mandated treatment of cider sufficient for a 5-log reduction of the target pathogen. Pasteurization has been suggested as the treatment most likely to achieve a 5-log reduction, with Escherichia coli O157:H7 as the target pathogen. Regulators and processors need a reliable method for verifying pasteurization, and apple cider polyphenol oxidase (PPO) activity was studied as a potential intrinsic index for thermal pasteurization. The effect of pasteurization conditions and apple cider properties on PPO activity and survival of three pathogens (E. coli O157:H7, Salmonella, and Listeria monocytogenes) was studied using a Box-Behnken response surface design. Factors considered in the design were pasteurization conditions, i.e., hold temperature (60, 68, and 76 degrees C), preheat time (10, 20, 30 s), and hold time (0, 15, 30 s), pH, and sugar content ((o)Brix) of apple cider. Response surface contour plots were constructed to illustrate the effect of these factors on PPO activity and pathogen survival. Reduction in PPO activity of at least 50% was equivalent to a 5-log reduction in E. coli O157:H7 or L. monocytogenes for cider at pH 3.7 and 12.5 (o)Brix. Further studies, however, are needed to verify the relationship between PPO activity and pathogen reduction in cider with various pH and (o)Brix values.

Beverages↗

Modified immunoliposome sandwich assay for the detection of Escherichia coli O157:H7 in apple cider.

Detection of Escherichia coli O157:H7 in fruit juices such as apple cider is necessary for diagnosis of infection and epidemiological investigations. However, inhibitors in the apple cider, such as endogenous polyphenols and acids, often decrease the sensitivity of PCR assays and immunoassays, thus routinely requiring laborious cell separation steps to increase the sensitivity. In the current study, polyethylene glycol (PEG)-derivatized liposomes encapsulating sulforhodamine B were tagged with anti-E. coli O157:H7 antibodies and used in an immunoliposome sandwich assay for the detection of E. coli O157:H7 in apple cider. Even without prior separation, this assay can detect E. coli O157:H7 in apple cider samples inoculated with as few as 1 CFU/ml after an 8-h enrichment period. The lower limit of detection in pure cultures without enrichment was 7 x 10(3) CFU/ml (280 CFU/40-microl sample). PEGylated immunoliposomes are suitable as an analytical reagent for the detection of E. coli O157:H7 in fruit juices containing polyphenols.

Antibodies, Bacterial↗

Power ultrasound treatment of Listeria monocytogenes in apple cider.

Inactivation experiments with Listeria monocytogenes 10403S, an ultrasound-resistant strain, were conducted at sublethal (20, 30, and 40 degrees C) and lethal (50, 55, and 60 degrees C) temperatures in saline solution (pH 7.0), acidified saline solution (pH 3.4), and apple cider (pH 3.4) with and without application of ultrasound (20 kHz, 457 mW.ml(-l)). The survival of recoverable L. monocytogenes 10403S in apple cider was evaluated, and the effects of temperature, ultrasound, pH, and food matrix on inactivation were studied. Application of ultrasound increased the inactivation rate at both sublethal and lethal temperatures. Additional death of L. monocytogenes 10403S was due to low acidity at the lethal temperatures. The reduction in surviving L. monocytogenes 10403S followed first order kinetics at sublethal temperatures, but at lethal temperatures, a two-section linear model described the inactivation behavior. The bactericidal effect of thermosonication was additive in apple cider. The survival tests of L. monocytogenes 10403S in apple cider indicated the possibility of using a mild treatment condition in combination with ultrasound to achieve a 5-log reduction in number of listerial cells.

Beverages↗

Pathogen reduction in unpasteurized apple cider: adding cranberry juice to enhance the lethality of warm hold and freeze-thaw steps.

U.S. Food and Drug Administration (FDA) regulations require processors of apple cider sold wholesale to use processing steps that ensure a 5-log reduction in numbers of the pertinent pathogen, generally considered to be Escherichia coli O157:H7. Current widely used validated pathogen-reduction steps are thermal pasteurization and UV light treatment. These techniques may be unaffordable or undesirable for some processors. This study investigated the cran-cider process, which is the addition of cranberry juice at a 15% (vol/vol) level, followed by warm hold (45 degrees C for 2 h) and freeze-thaw steps (-20 degrees C for 24 h, 5 degrees C for 24 h). When enumeration procedures did not include injury repair, the cran-cider process achieved a > or = 5-log reduction in numbers of E. coli O157:H7, Salmonella serovars, and Listeria monocytogenes. However, an injury-repair step was included in the pathogen enumeration procedure in confirmatory trials, and the resulting E. coli O157:H7 reductions of 3.5 to 4.2 log did not meet the FDA requirement. Consumer evaluation of apple cider subjected to the cran-cider process was favorable with a mean (n = 197) score of 5.8 on a seven-point hedonic scale (where 6 equals "like moderately") and 89% of panelists giving the product a positive score of 5, 6, or 7. The cran-cider process provides a novel way to improve microbial safety of unpasteurized apple cider, but it does not meet FDA-mandated pathogen reductions for wholesalers. However, cider makers selling apple cider only at retail could use the process to improve the safety of their product, provided containers were labeled with the FDA-mandated consumer warning.

Beverages↗