JOURNAL OF FOOD SCIENCE

MICROBIOLOGY

Control of Listeria monocytogenes in

Vacuum-Packaged Pre-Peeled Potatoes
VIJAY K. JUNEJA, STEFAN T. MARTIN, and GERALD M. SAPERS

ABSTRACT for inoculum preparation prior to all experi-

Growth of Listeria monocytogenes on the surface of fresh peeled potatoes, treated ments.
with sulfite or a commercial browning inhibitor (CBI), packaged under vacuum
and stored at 4, 15 and 28°C was determined. At 4°C, L. monocytogenes did not Preparation of inoculum
grow in all treated potatoes even after 21 days. At 15°C, L. monocytogenes grew Inoculated BHI was incubated at 37°C for
to 7 log10 CFU/g within 12 days in the potatoes treated with sulfite or CBI. At 28°C, 18h. The cells were harvested by centrifuga-
L. monocytogenes population was greater than 3 log10 CFU/g by 24 h in all samples tion at room temperature for 10 min at 7,700
regardless of treatment. Sulfites or a CBI appeared to provide a measure of safety × g, the cell pellet washed twice, resuspend-
in pre-peeled potatoes packaged under vacuum when kept at proper refrigera- ed and diluted in sterile 0.1% peptone water
tion temperatures. (w/v) (Difco).
Key Words: Listeria monocytogenes, browning inhibitor, potatoes
Sample preparation and inoculation
Fresh, raw Russet potatoes ( 80-110g
each) were obtained from a commercial po-
INTRODUCTION life of pre-peeled potatoes, reduction of ox- tato distributor. Potatoes were abrasion-
L ISTERIA MONOCYTOGENES IS A FOOD - ygen levels at the surface by packaging un- peeled for 2 min with a Vegetable Peeler
borne pathogen of great concern to the food der vacuum or modified atmosphere, and (Model A1-15, Toledo Scale Co., Toledo,
industry because its ubiquitous occurrence in storage at refrigeration temperatures, are nec- OH) and held under water up to 30 min prior
the environment can lead to its presence in essary (Langdon, 1987; O’Beirne and Bal- to treatment. Potatoes received one of 4 dif-
foods (Armstrong, 1985; Brackett, 1988). lantyne, 1987; O’Beirne, 1988). As a facul- ferent treatments: (1) immersion in 1.2% (w/
Published studies have examined the inci- tative anaerobe, L. monocytogenes can thrive v) sodium bisulfite for 2 min, (2) immersion
dence and fate of L. monocytogenes in fresh in an environment with little oxygen, such in a 2% (w/v) solution of a commercial
vegetables (Beuchat, 1996; Petran et al., as that of a vacuum package. browning inhibitor (CBI) consisting of cit-
1988). In two surveys of fresh market pro- While antimicrobial activity of sulfites in ric acid, ascorbic acid, sodium acid pyrophos-
duce, L. monocytogenes was detected on many foods is well documented (Wedzicha, phate and L-cysteine HCl for 5 min, (3) im-
27.1% (Heisick et al., 1989a) and 25.8% (He- 1981), the activity of sulfite alternatives is mersion in a 2% solution of the CBI for 5
isick et al., 1989b) of potato samples tested. not known. Accordingly, our study was un- min, followed by a brief rinse with tap water
The organism has been isolated from the fe- dertaken to examine the fate of L. monocy- (30 mL rinse/kg potatoes), or (4) untreated
ces of animals (Weis and Seeliger, 1975) togenes on the surface of fresh peeled pota- control. Residual sulfite levels were deter-
which may be a potential source of fresh pro- toes, treated with sulfite or a commercial mined in uninoculated control samples by the
duce contamination (Schlech et al., 1983). sulfite alternative, packaged under vacuum Optimized Monier-Williams Method (Hill-
Methods of controlling enzymatic brown- and stored at 4°, 15° and 28°C. Our objec- ary et al., 1989).
ing in minimally processed produce involve tive was to correlate the pathogen levels with Duplicate sets of two potatoes ( 100g
chemical treatment with browning inhibitors. background microflora (aerobic and anaero- each) were weighed into filter stomacher bags
The most widely used browning inhibitor for bic), and to determine whether treatment of (SFB-0410; Spiral Biotech., Bethesda, MD)
fresh, pre-peeled potatoes is sulfite. Over 1 fresh, vacuum-packaged pre-peeled potatoes and inoculated with 1 mL of an appropriate
million people in the U.S. are sensitive to with sulfiting agents or a commercial brown- dilution of L. monocytogenes cell suspension
sulfites (FDA, 1987), but the use of sulfites ing inhibitor would inhibit growth of to yield a final concentration of 1-2 log10
on pre-peeled potatoes is still authorized. L. monocytogenes. CFU/g. Thereafter, the bags were manually
However, the possibility of future regulato- massaged to ensure even distribution of or-
ry action against the use of sulfites and a de- MATERIALS & METHODS ganisms on the surface of the potatoes. The
crease in consumer acceptance of sulfite- bags were placed in 17.8 × 20.3 cm barrier
treated products (McEvily et al., 1992) have Test organism bags (Koch Model 01 46 09, Kansas City,
increased the a need for alternative brown- Listeria monocytogenes strains Scott A MO). The oxygen transmission rate (by man-
ing inhibitors. Several alternatives have been (serotype 4b; clinical isolate from Food and ufacturer) of the nylon/polyethylene film was
developed including combinations of organ- Drug Administration, Cincinnati, OH) and 54.2 cc/m2 in 24h measured at 24°C and 75%
ic acids, phosphates, preservatives, and oth- Murray B (serotype 4a; clinical isolate from relative humidity. The bags were evacuated
er adjuncts (Duxbury, 1987; Langdon, 1987; Centers for Disease Control, Atlanta, GA) to a negative pressure of 1000 millibars and
Santerre et al., 1991). To extend the shelf- from our in-house culture collection, main- heat sealed using a Multivac Model A300/
tained in 15% glycerol at -80°C, were used. 16 gas packaging machine (Germany). Two
Authors Juneja and Sapers are with the U.S. Depart-
Cells were grown on tryptic soy agar (Difco replications were performed for each treat-
ment of Agriculture, Agricultural Research Service, Laboratories, Detroit, MI) slants at 35°C for ment. For each replicate experiment, two
Eastern Regional Research Center, 600 E. Mermaid 24h and stored at 4°C. Cultures were trans- bags of each treatment were prepared for each
Lane, Wyndmoor, PA 19038 and author Martin is with ferred periodically to maintain viability and sampling time and temperature. Controls
EPL Technologies, Inc., 2 International Plaza, Suite 245,
Philadelphia, PA 19113-1507. used as stock cultures. Brain Heart Infusion consisted of inoculated, untreated potatoes
broth (BHI; Difco) was used to grow cultures stored under vacuum.

Volume 63, No. 5, 1998—JOURNAL OF FOOD SCIENCE 911

Growth of Listeria Monocytogenes In Vacuum-packaged Potatoes . . .

Storage and sampling Table 1—Growth of background bacterial populationsa as affected by treatments in vacuum-
packaged pre-peeled potatoes stored at 4°C
The inoculated vacuum-packaged potato
samples were stored at 4°, 15° and 28°C. Treatment Day
Samples stored at 4°C were analyzed on days 0 8 14 21
0, 8, 14, and 21. Samples stored at 15°C were Aerobic Sulfite 2.82 ± 0.15 6.36 ± 0.31 6.31 ± 0.36 6.27 ± 0.28
analyzed on days 0, 3, 6, 9, and 12; and those Plate CBI 2.82 ± 0.15 6.29 ± 0.26 6.92 ± 0.27 7.74 ± 0.14
stored at 28°C were analyzed at 0, 4, 8, 12, Counts CBI-RINSE 2.82 ± 0.15 6.27 ± 0.37b 6.22 ± 0.14b 6.13 ± 0.25b
Control 2.82 ± 0.15 6.46 ± 0.21 7.65 ± 0.37b 9.60 ± 0.28b
18, 24, 48, and 72h. Samples were observed
for visual evidence of spoilage or presence Anaerobic Sulfite 2.69 ± 0.22 6.74 ± 0.15 6.47 ± 0.28 5.53 ± 0.28
of off-odor. Plate CBI 2.69 ± 0.22 6.29 ± 0.40 5.67 ± 0.16 7.54 ± 0.17
Counts CBI-RINSE 2.69 ± 0.22 7.83 ± 0.26 6.40 ± 0.36 5.99 ± 0.33
Control 2.69 ± 0.22 7.46 ± 0.36 8.28 ± 0.28 8.91 ± 0.32
Bacterial enumeration
At the scheduled sampling time, sterile CVT Sulfite 2.47 ± 0.19 7.29 ± 0.23 8.32 ± 0.43 5.25 ± 0.16
0.1% peptone water (w/v) was added to each Plate CBI 2.47 ± 0.19 6.48 ± 0.23 7.34 ± 0.26 7.68 ± 0.25
Counts CBI-RINSE 2.47 ± 0.19 7.41 ± 0.15 7.75 ± 0.33 7.95 ± 0.27
bag to give a 1:1 (w/v) dilution and the con- Control 2.47 ± 0.19 7.31 ± 0.27 7.86 ± 0.14 8.89 ± 0.32
tents were homogenized for 1 min with a a All plate counts are log
10 CFU/g and represent means ± standard deviations of two replications.
stomacher Lab-blender (Model 400, Spiral b Samples exhibited evidence of spoilage (undesirable odor and appearance).

Systems, Inc). Further serial dilutions of each

sample were made using the same diluent as
required. This was followed by spiral plat-
ing (Spiral Systems Model D plating instru-
ments; Cincinnati, OH) of each dilution in
duplicate onto modified Modified Vogal
Johnson agar (MMVJ), Tryptic Soy agar
(TSA; Difco), or Crystal Violet Tetrazolium
agar (CVT; Difco) dishes. Also, 0.1 or 0.5
mL of undiluted suspension was surface plat-
ed, where relevant. The MMVJ agar plates
were incubated for 48h at 37°C for L. mono-
cytogenes enumeration, and CVT agar plates
were incubated at 28°C for 24h for psy-
chrotrophic Gram- negative bacterial counts.
The “total” aerobic and anaerobic colony
forming units (cfu) were determined after
aerobic and anaerobic incubation of TSA
plates at 37°C for 1 and 2 days, respectively.
RESULTS & DISCUSSION
AT 4°C, L. MONOCYTOGENES GREW RAPID-
ly in controls to almost 4 log10 CFU/g as early
as Day 8, but the organism did not grow at
that temperature in all samples treated with
sulfite (residual sulfite levels 237 ppm ± 55
ppm) or the Commercial Browning Inhibi-
tor (CBI), with or without a post-treatment
rinse, in 21 days (Fig. 1). The population
densities of aerobic, anaerobic and psy-
chrotrophic bacteria in treated and control
samples were greater than 6 log10 CFU/g by
Day 8 (Table 1). However, only samples
treated with the CBI followed by a rinse
showed marked evidence of spoilage (unde-
sirable odor and appearance) on this day of
observation. Hence, lacking or showing only
slight indications of spoilage, the potential-
ly hazardous controls might be perceived to
be edible on day 8. The levels of naturally
occurring microflora were consistently high
in all samples at the end of the 21-day stor-
age; all samples except those treated with
sulfite and the CBI (unrinsed) exhibited evi-
dence of spoilage. At 15°C, L. monocytoge-
nes rapidly grew to >6 log10 CFU/g in the
controls by day 3 (Fig. 2). The aerobic, anaer-
obic, and gram-negative psychrotroph lev-
els were 8 log10 CFU/g and there was visual
evidence of spoilage at day 3 (Table 2). L.
monocytogenes grew in all treated samples,
but growth was slower than in controls. By
day 3, growth of the pathogen had not ex-
ceeded 4.5 log10 CFU/g (only 2.5 log10 CFU/
g increase) in any of the treated potato sam-
ples. While the aerobic, anaerobic and psy-
chrotrophic bacteria plate counts were >6
log10 CFU/g, all treated potato samples were
considered to be of normal odor and color.
On day 6, L. monocytogenes counts were >5
log10 CFU/g in all samples, and the levels of
aerobic, anaerobic and Gram-negative psy-
chrotrophic bacteria were >8 log10 CFU/g.
However, only controls and those samples
treated with the CBI followed by a rinse had
developed spoilage odors and appearance.
Neither off odor nor discoloration was noted
in samples treated with sulfite or the CBI
(unrinsed) by Day 12, although though
growth of the pathogen exceeded 7 log10
CFU/g and background bacterial populations
were extremely high. Hence, use of sulfite
or the CBI in combination with vacuum pack-
aging created the same potentially hazard-
ous situation when held at the abusive tem-
perature of 15°C, as found in controls at 4°C.
At 28°C, L. monocytogenes levels were
below the limit of detection ( 1 CFU/g) in all
treated samples by 12h (Fig. 3). In contrast,
the population density increased in the con-
trol potatoes to 5 log10 CFU/g within 12h.

Fig. 1—Growth of L. monocytogenes in Fig. 2—Growth of L. monocytogenes in Fig. 3—Growth of L. monocytogenes in

treated vacuum-packaged pre-peeled treated vacuum-packaged pre-peeled treated vacuum-packaged pre-peeled
potatoes stored at 4°C. Log10 CFU/g of potatoes stored at 15°C. potatoes stored at 28°C. Log10 CFU/g of
the organism below the limit of detec- the organism below the limit of detec-
tion ( 1 CFU/g) were considered at 0. tion ( 1 CFU/g) were considered at 0.

912 JOURNAL OF FOOD SCIENCE—Volume 63, No. 5, 1998

Growth of Listeria Monocytogenes In Vacuum-packaged Potatoes . . .
Table 2—Growth of background bacterial populationsa as affected by treatments in vacuum- abuse, especially since the product might still
packaged pre-peeled potatoes stored at 15°C
exhibit normal odor, color, and texture. Tem-
Day perature abuse could occur during transpor-
Treatment 0 3 6 9 12 tation, distribution, storage or handling in
Aerobic Sulfite 2.82 ± 0.22 5.92 ± 0.24 9.59 ± 0.48 10.37 ± 0.28 10.41 ± 0.32
supermarkets or by consumers, primarily due
Plate CBI 2.82 ± 0.22 7.02 ± 0.31 9.54 ± 0.51 10.54 ± 0.33 9.42 ± 0.20 to equipment malfunction or electrical out-
Counts CBI-RINSE 2.82 ± 0.22 6.89 ± 0.32 10.15 ± 0.33b 11.16 ± 0.17b 9.35 ± 0.38b age.
Control 2.82 ± 0.22 8.25 ± 0.13 b 10.08 ± 0.28b 11.29 ± 0.28b 9.81 ± 0.13b Institutional foodservice organizations,
Anaerobic Sulfite 2.69 ± 0.17 5.99 ± 0.12 8.38 ± 0.32 9.22 ± 0.16 9.50 ± 0.13 catering establishments and retail consum-
Plate CBI 2.69 ± 0.17 6.83 ± 0.37 9.60 ± 0.22 10.99 ± 0.14 9.44 ± 0.57 ers who demand high quality, convenient
Counts CBI-RINSE 2.69 ± 0.17 7.87 ± 0.26 9.73 ± 0.29 10.68 ± 0.12 9.41 ± 0.17 foods may be consumers of vacuum-pack-
Control 2.69 ± 0.17 7.94 ± 0.31 10.30 ± 0.13 11.14 ± 0.32 10.34 ± 0.11
aged, pre-peeled potatoes. Consumers are
CVT Sulfite 2.47 ± 0.13 7.15 ± 0.32 10.22 ± 0.28 10.36 ± 0.41 9.77 ± 0.27 most likely to accept or reject such a product
Plate CBI 2.47 ± 013 6.51 ± 0.35 9.50 ± 0.26 11.04 ± 0.42 8.70 ± 0.16 based primarily on appearance and odor.
Counts CBI-RINSE 2.47 ± 0.13 6.89 ± 0.41 9.82 ± 0.13 10.87 ± 0.26 8.59 ± 0.31
Control 2.47 ± 0.13 7.87 ± 0.16 9.35 ± 0.11 10.47 ± 0.19 9.74 ± 0.32 Results indicate that potentially hazardous
a All plate counts are log
situations could occur in vacuum-packaged
10 CFU/g and represent means ± standard deviations of two replications.
b Samples exhibited evidence of spoilage (undesirable odor and appearance). and temperature abused potatoes in the ab-
sence of apparent spoilage signs. While the
cooking procedures for raw pre-peeled pota-
toes would be expected to inactivate
The population was greater than 5 log10 CFU/ When pre-peeled potatoes were dipped in L. monocytogenes, high levels of this patho-
g in samples treated with CBI (unrinsed and a solution of 2% ascorbic acid and 1% citric gen on raw product present the risk of cross-
rinsed) at 18h, although these samples did acid prior to vacuum-packaging and cook- contamination with other foods during han-
not appear to be spoiled. Initial levels of aer- ing, growth and toxin production of pro- dling. Food processors should be aware that
obic, anaerobic, and gram-negative psy- teolytic Clostridium botulinum type B was treatments in addition to vacuum-packaging
chrotrophic bacteria were >3 log10 CFU/g, inhibited for 70 days at 15°C and at least 28 are needed to control L. monocytogenes in
and ranged from 4 to 10 log10 CFU/g at 24h days at 20°C compared to controls which pre-peeled potatoes. The CBI we tested (both
(Table 3). All samples except those treated were toxic after 60 and 14 days at 15 and with and without a post-treatment rinse) pro-
with sulfite appeared to be spoiled by 24h. 20°C, respectively (Notermans et al. 1985). vided a similar level of safety (at 4 and 15°C)
Note that L. monocytogenes grew (3 log10 In our results, treatment with sulfites and the for L. monocytogenes control as the sulfite
CFU/g by 24h) in sulfite-treated potatoes that CBI (with or without a rinse) reduced growth treatment. It may therefore be considered as
were perceived to be unspoiled. of L. monocytogenes compared to controls a replacement for sulfites, particularly if
The antimicrobial effects of sulfites, as at all storage temperatures. Complete inhi- employed in conjunction with refrigeration.
well as that of the citric acid found in the bition for the duration of storage was ob- However, sufficient evidence exists to docu-
CBI formulation, are well-known (Davidson served in the treated samples at 4°C. There- ment temperature abuse as a common occur-
and Juneja, 1989). The most important fac- fore, samples treated with sulfites or the CBI rence at both the retail and consumer levels.
tor affecting the antimicrobial activity of (with or without a rinse) are unlikely to have Manufacturers should assume that tempera-
sulfites is pH. Increased effectiveness at low dangerous levels of L. monocytogenes if the ture abuse will probably occur at some point
pH is likely due to the ability of un-ionized product at low contamination levels is kept during the distribution of a refrigerated food
sulfur dioxide to pass across the cell mem- under low refrigeration temperatures. At product (NFPA, 1988). Surveys of retail food
brane (Rose and Pilkington, 1989). Regard- 15°C, however, L. monocytogenes grew >3 stores and consumer refrigeration units have
ing citric acid, the mechanism of inhibition log10 CFU/g in all samples and as high as 7 revealed that holding temperatures of >10°C
by citrate has been hypothesized to be relat- log10 CFU/g in some samples before obvi- are common (Anonymous, 1989; Bryan et
ed to its ability to chelate metal ions, in ad- ous product spoilage. Therefore, the patho- al., 1978; Daniels, 1991; van Grade and
dition to its pH-lowering effects (Davidson gen could present a danger if this product Woodburn, 1987; Wyatt and Guy, 1980).
and Juneja, 1989). received prolonged, extreme temperature Therefore, it is unrealistic to rely on refrig-

Table 3—Growth of background bacterial populations a as affected by treatments in vacuum-packaged pre-peeled potatoes stored at 28°C
Time (h)
Treatment 0 4 8 12 18 24 48
Aerobic Sulfite 4.22 ± 0.09 2.98 ± 0.11 3.65 ± 0.46 2.76 ± 0.28 3.77 ± 0.33 6.53 ± 0.19 8.37 ± 0.53b
Plate CBI 4.22 ± 0.09 2.86 ± 0.24 3.03 ± 0.37 5.38 ± 0.28 6.17 ± 0.19 8.54 ± 0.14b 9.03 ± 0.27b
Counts CBI-RINSE 4.22 ± 0.09 2.79 ± 0.35 3.32 ± 0.26 5.08 ± 0.42 5.32 ± 0.27 7.26 ± 0.21b 8.32 ± 0.16b
Control 4.22 ± 0.09 3.71 ± 0.26 4.29 ± 0.26 7.12 ± 0.05 7.80 ± 0.08 10.35 ± 0.19b 8.82 ± 0.42b

Anaerobic Sulfite 3.36 ± 0.15 5.20 ± 0.28 5.41 ± 0.39 4.28 ± 0.43 3.44 ± 0.32 4.33 ± 0.59 7.28 ± 0.31
Plate CBI 3.36 ± 0.15 5.28 ± 0.17 4.94 ± 0.32 4.24 ± 0.21 6.12 ± 0.37 9.67 ± 0.13 8.83 ± 0.29
Counts CBI-RINSE 3.36 ± 0.15 3.74 ± 0.11 3.34 ± 0.13 4.45 ± 0.28 5.35 ± 0.33 6.13 ± 0.28 8.32 ± 0.44
Control 3.36 ± 0.15 5.22 ± 0.27 4.84 ± 0.37 7.15 ± 0.35 8.45 ± 0.06 8.93 ± 0.19 8.81 ± 0.43

CVT Sulfite 3.59 ± 0.12 3.54 ± 0.04 3.96 ± 0.11 3.91 ± 0.32 3.82 ± 0.19 5.12 ± 0.51 7.12 ± 0.23
Plate CBI 3.59 ± 0.12 3.48 ± 0.24 3.97 ± 0.26 4.12 ± 0.17 6.44 ± 0.36 7.33 ± 0.07 9.34 ± 0.43
Counts CBI-RINSE 3.59 ± 0.12 4.19 ± 0.08 3.47 ± 0.08 4.97 ± 0.33 4.66 ± 0.08 6.96 ± 0.33 7.85 ± 0.34
Control 3.59 ± 0.12 3.95 ± 0.31 3.87 ± 0.17 6.64 ± 0.31 7.30 ± 0.50 8.59 ± 0.53 8.61 ± 0.41
a All plate counts are log
10 CFU/g and represent means ± standard deviations of two replications.
b Samples exhibited evidence of spoilage (undesirable odor and appearance).

Volume 63, No. 5, 1998—JOURNAL OF FOOD SCIENCE 913

Growth of Listeria Monocytogenes In Vacuum-packaged Potatoes . . .
eration for the safety of pre-peeled potato
products.
Our results indicate that institutional us-
ers and retailers should rigorously control
storage temperatures. This product should not
be marketed without continuous temperature
control from production to consumption.
Time/temperature indicators on each pack-
age may be useful to indicate if temperature
abuse had occurred and whether a potential
hazard might exist.
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Mention of brand or firm names does not constitute an endorse-
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nature not mentioned.