DIPHENYLAMINE RESIDUES IN APPLES AND APPLE CIDER

Diphenylamine (DPA) is an antioxidant which is widely used on apples for scald control during fruit storage. Since appreciable residues of DPA remain on apples when sold, it was of interest to study the extent of possible transfer of the compound to cider. Cider was expressed from five cultivars ofDPA-treated apples following their storage and the original fruit, the pomace and cider was analyzed for DPA residues. Only traces of the compound were found in cider with virtually all of the DPA found concentrated in the pomace. The toxicologic significance of the results are discussed.     

Use of hazard analysis critical control point and alternative treatments in the production of apple cider.

The purpose of this study was to evaluate the practices of Maryland cider producers and determine whether implementing hazard analysis critical control point (HACCP) would reduce the microbial contamination of cider. Cider producers (n = 11) were surveyed to determine existing manufacturing practices and sanitation. A training program was then conducted to inform operators of safety issues, including contamination with Escherichia coli O157:H7, and teach HACCP concepts and principles, sanitation procedures, and good manufacturing practice (GMP). Although all operators used a control strategy from one of the model HACCP plans provided, only one developed a written HACCP plan. None developed specific GMP, sanitation standard operating procedures, or sanitation monitoring records. Six operators changed or added production controls, including the exclusion of windfall apples, sanitizing apples chemically and by hot dip, and cider treatment with UV light or pasteurization. Facility inspections indicated improved sanitation and hazard control but identified ongoing problems. Microbiological evaluation of bottled cider before and after training, in-line apples, pomace, cider, and inoculated apples was conducted. E. coli O157:H7, Salmonella, or Staphylococcus aureus were not found in samples of in-line apple, pomace, and cider, or bottled cider. Generic E. coli was not isolated on in-coming apples but was found in 4 of 32 (13%) in-line samples and 3 of 17 (18%) bottled fresh cider samples, suggesting that E. coli was introduced during in-plant processing. To produce pathogen-free cider, operators must strictly conform to GMP and sanitation procedures in addition to HACCP controls. Controls aimed at preventing or eliminating pathogens on source apples are critical but alone may not be sufficient for product safety.     

Prevalence of Escherichia coli in apple cider manufactured in Connecticut.

Cider samples obtained from 11 cider mills operating in Connecticut during the 1997 to 1998 production season were tested for the presence of Escherichia coli. Cider production began in mid August and continued through March, with peak production in September and October. Of 314 cider samples tested, 11 (4%) were found to contain E. coli. Of the 11 mills, 6 (55%) tested positive for E. coli in the cider at least once during the production year. E. coli was first observed in cider samples produced in mid to late October and was not detected in samples made after January. A trend was observed for cider to decrease in acidity and increase in Brix (soluble sugars) throughout the production season. No correlation between pH and soluble sugars of cider and the presence of E. coli was detected. Eight mills used both dropped apples and tree-picked apples, whereas three mills used tree-picked apples only. The use of dropped apples in cider production began 5 weeks before the first detection of E. coli in cider. E. coli was isolated from cider samples produced using dropped apples and from samples produced using only tree-picked apples. No direct correlation between the use of dropped apples or tree-picked apples and the presence of E. coli in the cider was observed. An association between the time of apple harvest and the appearance of E. coli in cider was noted. For mills providing adequate records, all contaminated cider was produced from apples harvested between mid October and mid November.     

高效液相色谱法测定苹果酒中的多酚物质

建立了苹果酒中7种多酚物质的分析检测方法。使用该方法，各个多酚物质分离时间短，分离效果好，且回收率均超过80％、用该办法对自制苹果酒中多酚物质进行检测，得出样品中均有：儿茶素、咖啡酸、香豆酸等7种多酚物质。     

Isolation and characterization of Escherichia coli recovered from Maryland apple cider and the cider production environment

Contaminated apple cider has been implicated in several Escherichia coli O157:H7 outbreaks. In an attempt to investigate sources and modes of entry of E. coli into apple cider, samples of fresh apple, pomace, and cider and equipment and mill floor swabs were analyzed for standard plate counts (SPC), total coliforms (TC), fecal coliforms (FC), and E. coli. E. coli was isolated from 14 (33%) of 42 samples of bottled fresh cider, from food equipment in 6 (67%) of 9 mills, and from apples, pomace, or cider in 7 (78%) of 9 mills. Seventy-five E. coli isolates were further characterized for Shiga toxin-producing E. coli (STEC)-associated virulence factors, antimicrobial susceptibility, and pulsed-field gel electrophoresis (PFGE) type. No E. coli O157:H7 or other STEC was identified. Serotyping and PFGE revealed 64 distinct profiles, suggesting that recovered E. coli arose from multiple independent sources. However, on one occasion, E. coli isolated from the source apple sample was closely related to the E. coli identified in the finished cider sample. E. coli isolates were further tested for antimicrobial susceptibility to 17 antimicrobial agents of human and veterinary importance. Fourteen (19%) of the 75 isolates were resistant to at least one of the antimicrobial agents tested, and 9 (12%) were resistant to at least two of these agents. Of the resistant isolates recovered, 64% were resistant to tetracycline and 57% were resistant to streptomycin. Overall, the level of E. coli contamination in source apple samples did not differ significantly from those in samples of pomace, cider at the press, and cider entering the bottling tank; therefore, source apples cannot be dismissed as a potential contributor of E. coli to the cider-making process.     

Apple quality,storage, and washing treatments affect patulin levels in apple cider

Patulin is a mycotoxin produced primarily by Penicillium expansum, a mold responsible for rot in apples and other fruits. The growth of this fungus and the production of patulin are common in fruit that has been damaged. However, patulin can be detected in visibly sound fruit. The purpose of this project was to determine how apple quality, storage, and washing treatments affect patulin levels in apple cider. Patulin was not detected in cider pressed from fresh tree-picked apples (seven cultivars) but was found at levels of 40.2 to 374 microg/liter in cider pressed from four cultivars of fresh ground-harvested (dropped) apples. Patulin was not detected in cider pressed from culled tree-picked apples stored for 4 to 6 weeks at 0 to 2 degrees C but was found at levels of 0.97 to 64.0 microg/liter in cider pressed from unculled fruit stored under the same conditions. Cider from controlled-atmosphere-stored apples that were culled before pressing contained 0 to 15.1 microg of patulin per liter, while cider made from unculled fruit contained 59.9 to 120.5 microg of patulin per liter. The washing of ground-harvested apples before pressing reduced patulin levels in cider by 10 to 100%, depending on the initial patulin levels and the type of wash solution used. These results indicate that patulin is a good indicator of the quality of the apples used to manufacture cider. The avoidance of ground-harvested apples and the careful culling of apples before pressing are good methods for reducing patulin levels in cider.     

Characterization of Apples and Apple Cider Produced by a Guelph Area Orchard

Thermal stability of food-borne pathogens in apple cider is influenced by the composition of the product. As a preliminary step to determine the effect of pasteurization of apple cider on the survival of Escherichia coli O157:H7, a study was carried out to characterize apples and unpasteurized apple cider produced by a Guelph area orchard. Samples of commercial unpasteurized cider and the constituent apples were collected over 13 wk from August to November 1998, and unpasteurized laboratory cider was made from the individual apple varieties. pH, titratable acidity, turbidity, total microbial counts, total solids and °Brix for filtered and unfiltered samples were measured. The maximum, minimum, and average values for all unpasteurized commercial cider samples were found as follows: pH, 3.71, 3.17, and 3.43; titratable acidity, 93.47, 49.46, and 69.95 mL of 0.1 N NaOH/100 mL; total solids, 13.21, 10.93, and 11.90%; °Brix, 13.01, 11.17, and 12.02; turbidity, 238.1, 145.1, and 204.9 nephelometric turbidity units; and total plate count, 4.91, 2.61, 3.75 log cfu/mL. There were no significant differences (P>0.05) between filtered and unfiltered samples. In addition, in commercial unpasteurized cider, there were no significant differences (P>0.05) with respect to any of the factors with the time of processing. The composition of the unpasteurized laboratory cider made from individual apple varieties was dependent on the variety, but was generally within the ranges from the published literature values. McIntosh apples showed a significant (P\le0.05) decrease in titratable acidity with time of harvest. The results suggest that it is necessary to take the composition of commercial apple cider into account when developing thermal inactivation models for food-borne pathogens.     

Characterization of Apples and Apple Cider Produced by a Guelph Area Orchard

Thermal stability of food-borne pathogens in apple cider is influenced by the composition of the product. As a preliminary step to determining the effect of pasteurization of apple cider on survival of E. coli O157:H7, a study was carried out to characterize apples and unpasteurized apple cider produced by a guelph area orchard. Samples of commercial unpasteurized cider and the constituent apples were collected over 13 wk from August to November 1998, and unpasteurized laboratory cider was made from the individual apple varieties. pH, titratable acidity (TA), turbidity, total microbial counts, total solids and °brix for filtered and unfiltered samples were measured. The maximum, minimum, and average values for all unpasteurized commercial cider samples were found to be: pH, 3.71, 3.17, and 3.43; TA, 93.47, 49.46, and 69.95 mL of 0.1 N NaOH/100 mL; total solids, 13.21, 10.93, and 11.90%; °brix, 13.01, 11.17, and 12.02; turbidity, 238.1, 145.1, and 204.9 NTU; and total plate count, 4.91, 2.61, 3.75 log cfu·mL⁻¹. There were no significnat differences (P>0.05) between filtered and unfiltered samples. In addition, in commercial unpasteurized cider, there were no significant differences (P>0.05) with respect to any of the factors with time of processing. The composition of the unpasteurized laboratory cider made form individual apple varieties was dependent on the variety, but was generally within the ranges from published literature values. Mclntosh apples showed a significant (P≤0.05) decrease in TA with time of harvest. The results suggest that it is necessary to take the composition of commercial apple cider into account when developing thermal inactivation models for food-borne pathogens.     

Potential sources of microbial contamination in unpasteurized apple cider

A study was conducted to identify possible sources of microbial contamination and to assess the effect of good cleaning and sanitation practices on the microbial quality and safety of unpasteurized apple cider. Raw unwashed apples, washed apples, cleaning water, fresh cider, and finished cider samples were collected from five Ontario producers over 4 months and microbiologically tested. Total coliforms were found in 31, 71 and 38% of the unwashed apple, water, and washed apple samples, respectively. Escherichia coli was found in 40% of the water samples from one producer alone. The washing step was identified as a potential source of contamination, possibly due to water in the dump tanks seldom being refreshed, and because scrubbers, spray nozzles, and conveyors were not properly cleaned and sanitized. Higher total coliform counts (P < 0.0001) and prevalence (P < 0.0001) in fresh cider compared with those in unwashed apples and washed apples indicated considerable microbial buildup along the process, possibly explained by the lack of appropriate equipment sanitation procedures. Results showed that producers who had better sanitary practices in place had lower (P < 0.001) total coliform prevalence than the rest of the producers. Overall results show that good sanitation procedures are associated with improved microbial quality of fresh cider in terms of total coliforms and that operators who pasteurize and/or UV treat their product should still be required to have a sound good manufacturing practices program in place to prevent recontamination. Cryptosporidium parvum, an important pathogen for this industry, was found in different sample types, including washed apples, water, and fresh and finished cider.     

Biogenic amines content in Spanish and French natural ciders: application of qPCR for quantitative detection of biogenic amine-producers

Biogenic amines (BA) are low molecular weight nitrogenous bases commonly found in fermented foods and beverages and their consumption can induce undesirable reactions. In this work, the BA content in natural cider from Spain and France was determined. Samples from commercially available cider or obtained during the elaboration process were analyzed. A different profile and BA concentration was observed depending on cider origin. qPCR tools developed for the quantitative detection of BA producers from cheese and wine were tested in the cider samples. A good connection between the BA content and the presence of BA-producing microorganisms was observed. Based on these tools, BA-producing bacteria were isolated from the analyzed cider samples, including new potential histamine- and putrescine-producing Lactobacillus paracollinoides strains.     

Verifying apple cider plant sanitation and hazard analysis critical control point programs: choice of indicator bacteria and testing methods.

The objectives of this study were (i) to evaluate the survival of coliforms, Escherichia coli, and enterococci in refrigerated apple cider; (ii) to develop simple and inexpensive presumptive methods for detection of these bacteria; (iii) to perform a field survey to determine the prevalence of these bacteria on apples and in apple cider; and (iv) based on our results, to recommend the most useful of these three indicator groups for use in verifying apple cider processing plant sanitation and hazard analysis critical control point (HACCP) programs. Eight of 10 coliform strains (5 E. coli, 1 Enterobacter aerogenes, and 2 Klebsiella spp.) inoculated into preservative-free apple cider (pH 3.4, 13.3(o) Brix) survived well at 4 degrees C for 6 days (< or = 3.0 log10 CFU/ml decrease). Of 21 enterococci strains (Enterococcus faecalis, E. faecium, and E. durans), only 2 E. durans and 3 E. faecium strains survived well. Simple broth-based colorimetric methods were developed that detected the presence of approximately 10 cells of coliforms or enterococci. In three field studies, samples of unwashed apples (drops and picked), washed apples, and freshly pressed cider were presumptively analyzed for total coliforms, E. coli, and enterococci using qualitative and/or quantitative methods. Drop apples were more likely than picked apples to be contaminated with E. coli (26.7% vs. 0%) and enterococci (20% vs. 0%). Washing had little effect on coliform populations and in one field study was associated with increased numbers. Total coliform populations in cider ranged from < 1 CFU/ml to > 738 most probable number/ml, depending on the enumeration method used and the sample origin. E. coli was not recovered from washed apples or cider, but enterococci were present on 13% of washed apple samples. The qualitative coliform method successfully detected these bacteria on apples and in cider. Based on its exclusively fecal origin, good survival in apple cider, and association with drop apples, we conclude that E. coli is the most useful organism for verifying apple cider sanitation and HACCP programs.     

啤酒糟苹果醋不同发酵阶段挥发性香气成分及抗氧化性变化分析

采用顶空固相微萃取（HS-SPME）结合气质联用（GC-MS）法测定啤酒糟苹果汁、苹果酒、苹果醋的挥发性成分,探究啤酒糟苹果醋不同发酵阶段挥发性香气成分及抗氧化性变化。结果表明,苹果醋不同发酵阶段样品共检测出50种挥发性化合物,其中酯类16种、醇类8种、醛类8种、酸类7种、酚类4种、其他类4种、酮类2种、醚类1种。啤酒糟苹果汁、苹果酒、苹果醋中分别共检测出12种、15种、24种挥发性物质。抗氧化活性试验结果表明,DPPH自由基的清除率顺序为啤酒糟苹果酒＞啤酒糟苹果汁＞啤酒糟苹果醋,Fe3+还原能力顺序为啤酒糟苹果汁＞啤酒糟苹果酒＞啤酒糟苹果醋。     

Microbial levels in Michigan apple cider and their association with manufacturing practices

In recent decades, apple cider has been implicated in a series of outbreaks of foodborne illness. The objective of this study was to determine the presence and concentrations of pathogenic and indicator microorganisms in apple cider processed in Michigan and to evaluate the impact of thermal pasteurization, UV light radiation, and implementation of hazard analysis critical control point (HACCP) plans on these microbes. Cider samples were obtained from Michigan mills between 1997 and 2004 and analyzed for Escherichia coli O157:H7, Salmonella, generic E. coli, total coliforms, and aerobic bacteria. Neither E. coli O157:H7 nor Salmonella were detected in any tested cider samples, suggesting a very low frequency of pathogens in Michigan apple cider. The persistent and relatively high frequency of generic E. coli observed in samples obtained in all years indicates a continued risk of pathogen contamination in Michigan apple cider, especially when it is untreated. The use of thermal pasteurization or UV light radiation and reported implementation of HACCP plans were associated with lower frequency and counts of generic E. coli, total coliforms, and aerobic microorganisms. However, the relatively high counts of indicator organisms in some cider samples that were claimed to be treated according to these pathogen reduction measures indicates that some processors had inadequate practices, facilities, or equipment for pathogen reduction or did not consistently or adequately apply practices or pathogen-reduction equipment in an effective manner.     

STORAGE QUALITY OF PASTEURIZED AND UV TREATED APPLE CIDER

Two studies were conducted to assess the effect of hot-fill pasteurization at 63C and UV irradiation at 14 mJ/cm² on the quality and shelf-life of apple cider packaged under controlled conditions with minimal packaging contamination, and under pilot plant conditions resembling commercial operations. The processed cider was stored at 7C for up to 14 weeks in the first study and 4 weeks in the second. Microbiological, chemical and sensory tests were conducted weekly on cider samples. There were no significant differences among the fresh processed ciders with regard to taste and preference. All treatments achieved a reasonable reduction in microbial counts, although hot-fill pasteurization provided longer shelf-life. There were significant changes in pH, titratable acidity, soluble solids and turbidity of samples during storage. Hot-fill at 63C is a comparable alternative to flash pasteurization at 71C for 6 s for the production of safe quality cider at small cider mills.     

Aromatic Profile of Ciders by Chemical Quantitative,Gas Chromatography‐Olfactometry,and Sensory Analysis

Nine samples of Asturias cider have been analyzed for volatile, olfactometric, and sensorial profiles. The aromatic composition was mainly constituted by fusel alcohols and ethyl esters. Among the minor volatile compounds, fatty acids, volatile phenols, and alcohols were the main components. The olfactometric analysis revealed the existence of 55 aromatic areas, exhibiting a wide range of intensities. Components like amyl alcohols, 2‐phenylethanol, ethyl esters such as 2‐methylbutyrate, hexanoate and octanoate, hexanoic and octanoic acids 2‐phenylethyl acetate, 4‐ethyl guaiacol, and 4‐ethyl phenol could be considered as being part of the structure of cider aroma. The extract dilution analysis of one extract identified 2 volatile phenols (4‐ethyl guaiacol and 4‐ethyl phenol) among the most powerful odorants in cider. These components gave significant correlations with the sensory attributes sweet, spicy, and lees.     

Duplex PCR Method for Rapid Detection of Zymomonas mobilis in Cider

A duplex PCR method was developed for Z. mobilis species detection. Two sets of primers were used; the first one targeted a 530 bp region of the 23S rRNA gene, the amplified fragment corresponding to an internal control, while the second targeted a 900 bp region spanning the 16S and 23S rRNA gene regions and was Z. mobilis specific. This second primer pair was designed by comparison with corresponding sequences of a Sphingomonas sp., a bacterium that is phylogenetically the closest to Z. mobilis, and Gluconobacter oxydans that is taxonomically the closest relative found in cider. Results showed that the method was able to specifically amplify the two targets in all studied strains of Z. mobilis while only one fragment, corresponding to the internal control, was amplified for G. oxydans and the tested lactic acid bacteria commonly found in cider. The method threshold was determined by contaminating cider samples containing natural yeast and lactic acid bacteria flora. Within a 24 h period, ciders containing 10² CFU/mL of Zymomonas were detected as positives. The rapidity and reliability of this duplex PCR for detection of Z. mobilis directly from cider will certainly prove to be useful in detecting, early on during the fermentation phase, the presence of this spoilage microorganism. It will also be of interest as a complementary test when using the HACCP approach in the cider and beer industries.     

Malolactic Fermentation as a Technique for the Deacidification of Hard Apple Cider

ABSTRACT:  Malolactic fermentation (MLF), the conversion of malate to lactate, is an important process leading to the deacidification of hard apple cider. MLF is dependent on the levels of inhibitory factors such as sulfur dioxide and ethanol. To assess the effect of these 2 factors on MLF, hard apple cider was produced from pasteurized, unfiltered apple cider ( Malus domestica  cvs Red Delicious, Golden Delicious, Braeburn, and Fuji). Apple cider was treated with 2 levels of sulfur dioxide (50 and 80 ppm) and then fermented using  Saccharomyces cerevisiae  montrachet. After the primary fermentation, 1 set of the samples remained unadjusted and 100% ethyl alcohol was used to adjust other sets of samples to 7%, 9%, or 11% (v/v) ethanol. Following the ethanol adjustment,  Oenococcus oeni  MCW was used to initiate the MLF in half of the samples. Cider parameters monitored throughout the fermentations included organic acid content, titratable acidity, pH, ethanol production, and sugar content. Since samples containing either sulfur dioxide level had similar sugar utilization rates and ethanol production it was concluded that sulfur dioxide had no effect on the primary fermentation. Sulfur dioxide content was shown to have an impact on MLF. There was no difference in the rate of malic acid consumption, but lactic acid production was faster in the 50-ppm sulfur dioxide samples. MLF was not inhibited by ethanol content.     

Changes in headspace volatile attributes of apple cider resulting from thermal processing and storage

One attribute that frequently reflects a product's organoleptic impact is its aroma. In research presented here, a Fox-3000 Electronic Nose was used to determine and compare the aromatic profiles of samples of apple cider subjected to various thermal treatments. Initial results have indicated that exposure to temperatures of up to 90 °C for approximately 28 s acts to stabilize the aromatic properties of apple cider over a 7-day period. In contrast, there are significant changes in the aromatic profiles of fresh apple cider having no such thermal treatment over the same time period. A comparison of samples thermally treated at 60, 70, and 80 °C to non-processed cider showed minimal statistical difference on the basis of similarity indices obtained after 24-h of storage of the samples at 4 °C. Treatment at 90 °C showed significant differences from the unprocessed samples in the same test sequence. After storage for 7-days, samples that were thermally treated exhibited minimal differences from each other, however, were quite different from the non-processed sample on the basis of similarity index analysis. GC analysis was done to confirm potential differences between the samples measured using the electronic nose. Sensory testing is required to determine whether these differences affect the overall quality perception by consumers.     

Preference Testing of Whiskey Sour Formulations: Magnitude Estimation Versus the 9-Point Hedonic

Consumer preferences for different whiskey sour formulations were determined by a consumer laboratory panel and by a home panel using two different sensory techniques: the magnitude estimation procedure and the nine-point hedonic method. Use of the magnitude estimation testing procedure resulted in more statistically significant differences in preference in both the home panel and the consumer laboratory panel. Many more significant differences in preference were expressed by the home panel than by the consumer lab panel. The standard whiskey sour formula scored very well in the preference tests in comparison to the formula adjusted samples. In only two of the comparisons were the adjusted samples preferred over the standard, the 25% dilution of the standard and the 35° proof samples.