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The antimicrobial effects of cranberry against Staphylococcus aureus

Poh Yng Lian, T Maseko, M Rhee and K Ng
Food Science and Technology International 2012 18: 179 originally published online 13 March 2012
DOI: 10.1177/1082013211415159

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Article

The antimicrobial effects of cranberry against

Staphylococcus aureus

Poh Yng Lian, T Maseko, M Rhee and K Ng

Abstract
The antimicrobial effects of the American cranberry (Vaccinium macrocarpon) on a major food-borne
pathogen, Staphylococcus aureus, were investigated using commercially obtained LakewoodÕ organic
cranberry juice and Ocean SprayÕ cranberry juice cocktail and four other berry fruit extracts (acai berry,
strawberry, raspberry, and blueberry). The results showed that cranberry is a potent antimicrobial against S.
aureus and the most potent among the berries studied. The order of percentage inhibition of bacterial growth
at the same concentration of phenolic materials as gallic acid equivalents was Lakewood cranberry
juice > Ocean Spray cranberry juice  blueberry > acai berry  raspberry  strawberry. The antimicrobial
effect was not due to the acidity of the berries as NaOH-neutralized samples were almost as effective in
terms of percentage inhibition of viable cell growth. Solid-phase extraction of cranberry juice using C18 solid
phase showed that the antimicrobial effects reside exclusively with the C18-bound materials.

Keywords
Cranberry, antimicrobials, Staphylococcus aureus, phenolic phytochemicals
Date received: 2 March 2011; revised: 26 May 2011

INTRODUCTION by the production of various types of toxins (Kwon

Staphylococcus aureus is a Gram-positive coccus and
one of the major causes of food-borne illness through-
out the world (Asperger and Zangerl, 2002). The organ-
ism is usually found in large numbers in the nails, skin,
and hair of human beings where they find their way
into the foods by hands, coughing, and sneezing.
Unhygienic human handling is the most important
route of food contamination due to oversight of inci-
dences like skin lesions and infected wounds (Asperger
and Zangerl, 2002). The clinical condition of staphylo-
coccal food poisoning is identified as vomiting, some-
times accompanied by gastroenteritis (Bania et al.,
2006). This is caused by the ingestion of food that has
been contaminated by S. aureus (Dingers et al., 2000).
Staphylococcus aureus is also responsible for numerous
other diseases ranging from skin to soft tissue infections
et al., 2007).
The prevalence of antibiotic resistance by S. aureus is
the major reason for treatment failure as the ability for
resistance is easily transferred to other bacteria via a
range of gene transfer mechanisms (Kwon et al., 2007).
From a food safety perspective, the addition of chem-
ical preservatives is a common practice to enhance food
safety. However, consumers are becoming increasingly
wary of the side effects of chemical preservatives, which
lead to the increase in demand for natural preservatives
(Wu et al., 2008).
Plants are the primary source of a wide range of
naturally occurring phytochemicals that include
phenolic compounds that were formed as secondary
metabolite products (Zafra-Stone et al., 2007).

Department of Agriculture and Food Systems, Melbourne School

Food Science and Technology International 18(2) 179–186
of Land and Environment, University of Melbourne, Australia
! The Author(s) 2012 Reprints and permissions: Corresponding author:
sagepub.co.uk/journalsPermissions.nav K Ng, Department of Agriculture and Food Systems, Melbourne
DOI: 10.1177/1082013211415159 School of Land and Environment, University of Melbourne, Royal
fst.sagepub.com Parade, Parkville, Victoria 3010, Australia
Email: ngkf@unimelb.edu.au

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Food Science and Technology International 18(2)
Phenolic phytochemicals serve primarily as protection
in plants against free radicals and radiation damage
(Stevenson and Hurst, 2007). Dark-colored berry such
as cranberry represents one of the most important diet-
ary sources of phenolic compounds that can be categor-
ized into four main groups: simple phenolic acids, such
as gallic acid and caffeic acid, lignin fragments and
monomeric flavonoids (catechins, flavonols, and antho-
cyanins) and polymeric (proanthocyanidins; PCA)
flavonoids (Fang et al., 2009; Singh et al., 2009).
The American cranberry fruit, Vaccinium macrocar-
pon, is a traditional fruit originating from America and
was historically used by Native Americans as a health
promoting fruit (Apostolidis et al., 2008). Cranberry
has been shown to possess potent antioxidant activity
and antimicrobial properties linked to its phenolic
contents (Guo et al., 2007; Lacombe et al., 2010;
Nohynek et al., 2006). Cranberry, like most other
berries, contains B-type PAC, which is a short-chain
polymer of anthocyanidin units containing multiples
of the basic structure linked by a single ether bond
(Dixon et al., 2005). However, cranberry also contains
a unique A-type PAC which bears a second ether
linkage through the chroman’s oxygen (Howell, 2007).
A-type PAC is linked to antimicrobial effect in prevent-
ing urinary tract infection by P-fimbriated Escherichia
coli (Howell, 2007; Johnson-White et al., 2006; Liu
et al., 2006, 2008). The inhibition mechanism appears
to be due to the ability of A-type PAC molecules
inhibiting the attachment of E. coli to the epithelial
cells lining of the urinary tract, preventing infection.
Like all bacterial infections, antibiotic resistance is a
major health threat and naturally occurring antibacter-
ial agents are highly sort after (Foxman et al., 2000).
Recently, cranberry extracts have also been shown to
display antimicrobial effects on S. aureus, Listeria
monocytogenes, and E. coli strain by as yet undermined
mechanism of killing bacteria or inhibiting growth
(Lacombe et al., 2010; Wu et al., 2009).
This article compares the antimicrobial effect of two
commercially available cranberry juice products against
S. aureus and compares the cranberry juices with the
extracts of acai berry, blueberry, raspberry, and straw-
berry fruits. We also provided evidence for the anti-
microbial effect of cranberry residing with its phenolic
materials.
MATERIALS AND METHODS
Materials
Ocean SprayÕ cranberry juice cocktail light (manufac-
turer stated ingredients: water, 27% cranberry juice (v/v
from concentrates)), natural flavors (not specified), arti-
ficial sweetener (sucralose)), LakewoodÕ pure organic
cranberry juice (manufacturer stated ingredient: made
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from 100% fruit juice with no added water or sugar)
and acai berry, blueberry, strawberry, and raspberry
fruits were obtained from a local supermarket. Sep-
Pak C18 cartridges were from Waters Corporation.
Folin–Ciocalteu ReagentÕ (2N) and gallic acid were
from Sigma–Aldrich Inc. (St. Louis, MO, USA). All
other common laboratory chemicals used were of ana-
lytical grade or better. Water used was MilliporeÕ deio-
nized water.
Bacterial culture. Staphylococcus aureus strain
ACTT25923 (Seattle) was kept under refrigeration at
4  C as stock culture and reactivated weekly to main-
tain viability. To reactivate the culture, a single isolated
colony was obtained from the stock culture plate and
inoculated into 9 mL of brain heart infusion (BHI)
broth and incubated at 37  C for 24 h. An inoculation
loop is then used to prepare a streak plate from the
incubated BHI and the plate incubated for another
24 h at 37  C before refrigeration as further stock
culture.
Culture media. Baird Parker agar (BPA) plates were
prepared for the purpose of enumeration of bacteria.
BPA was prepared following the protocol specified by
the manufacturer’s instructions. The media powder
used was purchased from Oxoid. The required
amount of powdered media was added to 1 L of dis-
tilled water in Schott bottles and boiled to insure that
the powder was well mixed and dissolved. The bottles
were then autoclaved at 121  C for 15 min and allowed
to cool to 50  C before adding 50 mL of egg yolk tel-
lurite emulsion. The bottle was gently swirled to insure
even distribution and then aseptically poured into ster-
ile Petri dishes. The plates were then refrigerated at 4  C
until further use.
BHI broth was also prepared by evenly mixing the
powdered broth purchased from Oxoid. The required
amount of powder to make up 1 L of broth was 37 g.
The mixture was well shaken to insure that the powder
had dissolved and mixed evenly. The broth (9 mL) was
then distributed into individual McCartney bottles with
a Zipette Classic bottle-top dispenser. The bottles were
placed in racks and autoclaved at 121  C for 15 min and
allowed to cool. The bottles were refrigerated at 4  C
until further use.
Peptone water at a concentration of 0.01% w/v was
prepared for the purpose of dilution. The powdered
media was also obtained from Oxoid. One gram of
powder was evenly dissolved in 1 L of distilled water.
The peptone water (9 mL) was then distributed into
individual McCartney bottles. The bottles were placed
in racks and autoclaved at 121  C for 15 min and
allowed to cool. The bottles were refrigerated at 4  C
until further use.

Sterilized berry samples. Acai berry, blueberry, rasp-
berry, and strawberry juices were prepared from whole
fruits by blending individual berry with a commercial
blender and centrifugation in a bench-top centrifuge at
5000 r/min for 60 min to obtain clear juice.
The original pH values of the various berry samples
were fairly acidic; Lakewood cranberry (1.19), Ocean
Spray cranberry (2.96), acai berry (4.32), blueberry
(3.34), strawberry (3.69), and raspberry (3.23).
Lakewood cranberry has the strongest acidity and
required 0.5 M of NaOH (final concentration in
juice) to neutralize. Ocean Spray cranberry required
0.15 M while the other berries required less than
0.1 M of NaOH to neutralize.
Original (acidic) and pH neutralized (pH 7) juices
were individually sterilized by passage through sterile
Acrodisc 25 mm filter with a 0.2 mm Supor membrane.
Methods
Partially purified cranberry. A sample of the Lakewood
cranberry juice was subjected to solid-phase extraction
using a 30 mL Sep-Pak C18 solid-phase column. The
juice (50 mL) was applied to the column and washed
with 10 volume of water before elution with 5
volume of 100% methanol to recover the intensely col-
ored bound materials. The straight through and water
wash were combined, pH adjusted to neutral with
NaOH, solution evaporated to dryness at 40  C under
reduced pressure, and the dried materials reconstituted
with 25 mL water (2 concentrated). The methanol
eluate was similarly dried without pH adjustment (solu-
tion was neutral) and materials also reconstituted with
25 mL water (2  concentrated). Both samples were
sterilized by passage through 0.2 mm membrane filter
and stored at 4  C before analysis.
Quantification of phenolic contents. Samples (berries
juices and cranberry C18 straight through and metha-
nol eluates) were analyzed for their total phenolic con-
tents using Folin–Ciocalteu Reagent and gallic acid as
standards as described by Slinkard and Singleton
(1977). One milliliter of sample or standard solution
was mixed with 1 mL of freshly diluted Folin–
Ciocalteu Reagent (1 : 10 dilutions with water; 0.2 N)
in a plastic tube and shaken vigorously. After 10 min,
3 mL of Na2CO3 solution (10%, w/v) was added and
the mixture incubated at 40  C for 15 min with intermit-
tent shaking. The absorbance was measured using a
UV/vis spectrophotometer (UV-1601 Shimadzu) at
765 nm. A range of varying amounts of the sample
(n ¼ 4) or gallic acid standard (n ¼ 5) were determined.
The concentration of phenolic materials as gallic acid
equivalent (GAE) was calculated according to the fol-
lowing linear equation that was obtained from a plot of
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Lian et al.
mean absorbance (AU) versus mass (mg) from the gallic
acid standard plot: AU ¼ 0.0088 mg (sample) þ 0.0146
(R2 ¼ 0.9973).
Antimicrobial effects of berry extracts. Staphylococcus
aureus was cultured by taking an isolated colony from a
refrigerated stock plate and inoculating it into 9 mL of
BHI. The broth was incubated at 37  C for 24 h. To
obtain the initial bacterial count, 1 mL of the cultured
BHI was used to conduct 6  1 : 10 serial dilutions using
peptone water. Duplicate 100 mL of the 106 diluted
sample was then plated on BPA using the spread
plate method. The duplicate plates were incubated at
37  C for 24 h and their bacterial colonies counted.
For the antimicrobial effect study, 1 mL of the bac-
terial culture in BHI was pipetted out into separate
McCartney bottles, plus 8 mL of fresh BHI and 1 mL
of berry/test sample (total volume 10 mL). The bottles
were then incubated at 37  C for 24 h and a 100 mL
sample was taken from each bottle and subjected to
5  1 : 10 serial dilutions with peptone water.
Duplicate 100 mL was taken from the 103, 104, and
105 diluted samples and were plated on separate BPA
plates. The plates were incubated at 37  C for 24 h and
bacterial colonies counted to obtain the 24 h antimicro-
bial effect. For 48 h antimicrobial effect, the bottles
were further incubated for another 24 h and duplicate
100 mL samples were again taken from each bottle for
plating and incubation on BPA for 24 h and colonies
counted.
Statistical analysis. Each determination was performed
in duplicate. The number of bacterial cells were
reported as log CFU/mL (CFU, colony forming unit)
and treatments were assigned for comparison. Analyses
of variance were performed on the mean and range of
the results using the SAS General Linear Model pro-
cedures with SAS 9.2 software (Statistical Analysis
System Institute, Inc., Cary, NC, USA). Differences
among treatments were examined for the level of sig-
nificance by least significant difference. Significance of
differences was defined as p < 0.05.
RESULTS AND DISCUSSION
Phenolic contents of berry samples
Table 1 presents the phenolic contents of the berry sam-
ples as determined by a colorimetric method using
Folin–Ciocalteu Reagent and expressed as GAE
(Slinkard and Singleton, 1977). It can be seen that the
Lakewood organic 100% cranberry concentrate con-
tained more than twice the amount of phenolic materials
than Ocean Spray cranberry light cocktail. This is due to
the Ocean Spray cranberry containing only 27% of juice.
The acai berry, blueberry, strawberry, and raspberry
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Food Science and Technology International 18(2)

fruit extracts also contained substantial amounts of
phenolic materials. The phenolic contents were not
related to the weight of fruits as we are only concerned
with relating the phenolic contents (as GAE) to their
antimicrobial effects on the viability of S. aureus.
A sample of the Lakewood cranberry concentrate
was fractionated by solid-phase extraction using a
C18 solid phase open column into straight-through
and methanol-eluated fractions. The fractions were
reconstituted to twice their original concentrations
(i.e., half the original volume applied to the column).
Table 1. The total phenolic content of juices made with
berries
Berry samples
Ocean Spray cranberry juice
Lakewood cranberry juice
Acai berry
Strawberry
Blueberry
Raspberry
Lakewood cranberry C18
straight-through fractiona
Lakewood cranberry C18
methanol-eluated fractiona
GAE: gallic acid equivalent and ND: none detected.
a
Extraction fractions reconstituted in half the volume of original
sample applied to C18 column.
Not surprisingly, all the phenolic materials were recov-
ered in the C18-bound methanol-eluated fraction
(Table 1) as they have affinity for the C18 solid phase.
Antimicrobial effects of Ocean Spray and
Lakewood cranberry juice on S. aureus
The antimicrobial effects of Ocean Spray and
Lakewood cranberry juice were tested against S.
aureus. Five different concentrations (in terms the phen-
olic materials added as mg GAE/mL) of the original
(acidic) and pH-neutralized (pH 7) juice were tested
and the bacterial culture incubated for 24 and 48 h per-
iods. The results (Table 2) showed that the percentage
inhibition of S. aureus growth, as measured by viable
mg GAE/mL cell counts, compared to control without added juice
was greater in Lakewood than the Ocean Spray cran-
867  1
berry juice at the same concentration of phenolic mater-
2305  11
ials for a concentration range 0–86.6 mg GAE/mL
1006  4 bacterial culture tested. Higher concentration
743  9 Lakewood cranberry juice at 173.1 mg GAE/mL only
2018  8 increases the percentage inhibition slightly, indicating
771  1 optimal concentration effect has been reached (the
ND Ocean Spray cranberry juice could only be added to
86.6 mg GAE/mL in the assay protocol).
4585  15 The inhibition percentage, or number of cell deaths,
only increased slightly between the 24 and 48 h incuba-
tion period for all concentrations (Table 2), indicating
that a 24 h incubation period would be sufficient for
the study. At 86.6 mg GAE of cranberry/mL bacteria

Table 2. The viable cell counts and percentage inhibition compared to control of S. aureus incubated with varying
concentrations of Ocean Spray and Lakewood cranberry juice for 24 and 48 h incubation periods at original and
neutral pH

Viable cell counts (log CFU/mL) Inhibition (%)

Original pH 7 Original pH 7

24 h 48 h 24 h 48 h 24 h 48 h 24 h 48 h

Ocean Spray
cranberry (mg GAE/mL)
0 8.24  0.02 a 8.19  0.03 a 8.26  0.02 a 8.21  0.01 a 0 0 0 0
17.3 8.12  0.03 b 8.05  0.03 b 8.17  0.02 b 8.10  0.02 b 1.5 1.7 1.1 1.3
43.3 7.42  0.01 c 7.42  0.02 c 7.45  0.01 c 7.38  0.03 c 10.0 11.8 9.8 10.1
86.6 7.29  0.01 d 7.11  0.02 d 7.33  0.02 d 7.22  0.01 d 11.5 13.2 11.3 12.1
Lakewood
cranberry (mg GAE/mL)
0 8.23  0.02 a 8.21  0.03 a 8.26  0.03 a 8.22  0.02 a 0 0 0 0
17.3 8.00  0.05 b 7.83  0.01 b 8.08  0.01 b 7.96  0.02 b 2.8 4.6 2.2 3.2
43.3 7.32  0.02 c 7.19  0.04 c 7.39  0.08 c 7.27  0.01 c 11.1 12.4 10.5 11.6
86.6 7.16  0.02 d 6.99  0.07 d 7.24  0.05 d 7.07  0.03 d 13.0 14.9 12.3 13.9
173.1 7.06  0.02 e 6.86  0.01 e 7.12  0.01 e 6.95  0.02 e 14.2 16.4 13.8 15.4
CFU: colony forming unit and GAE: gallic acid equivalent.
Mean values followed by different letters within a column are significantly different (p < 0.05).

182
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 Lian et al.

culture, the inhibition achieved was 12.3–14.9% for
Lakewood and 11.5–13.2% for Ocean Spray cranberry
juice (Table 2). Both juice samples contain some ori-
ginal sugars attributed to the fruit source that might
contribute to promoting some degree of bacterial
growth. However, the flavoring and pectin additives
in the Ocean Spray cranberry juice might have compli-
cated further the antimicrobial effect by promoting
some bacterial growth that produces a lower percentage
inhibition observed, compared to the Lakewood cran-
berry juice which was made with 100% pure fruit juice
with no additives added.
Importantly, the results also showed that the anti-
microbial effects were not due to the strong acidity of
the cranberry juices as the percentage inhibitions were
similar for original and pH-neutralized juices across all
concentrations tested (Table 2). Early studies have
credited the antimicrobial effects of cranberry extracts
to the extreme acidity of the juice (Johnson-White
et al., 2006). However, it has since been demonstrated
by a couple of later studies (Lacombe et al., 2010; Wu
et al., 2008) that pH does not appear to play a central
role in the antimicrobial effects of berries. Our data
further supported this inference as the differences in
the overall inhibition of the pH-adjusted cranberry
treatments as compared to their original pH was only
less than 1.5%. The high acidity of cranberry juice is
believed to be due to the presence of organic acids (Van
Immerseel et al., 2006). In the case of the Lakewood
cranberry juice, the pH of the juice was very low, 1.19.
Antimicrobial effects of various berries on
S. aureus
In order to establish the effectiveness of cranberry as an
antimicrobial, an inhibition experiment was carried out
to compare both Lakewood and Ocean Spray cranberry
juices against four other berry juices prepared from
whole fruits (acai berry, blueberry, raspberry, and straw-
berry). The berry juices were tested at their neutralized
pH (7) and were administered at 86.6 mgGAE/mL bac-
terial culture for 24 and 48 h period incubation period.
The results (Table 3) showed that the order of antimicro-
bial activity against S aureus was Lakewood cranberry
juice > Ocean Spray cranberry juice  blueberry > acai
berry  raspberry  strawberry. Lakewood cranberry

Table 3. The viable cell counts and percentage inhibition (compared to control) of S. aureus for various pH-neutralized
berry samples at 86.6 mg GAE/mL bacterial culture for 24 and 48 h incubation periods

Viable cell counts (log CFU/mL) Inhibition (%)

Treatment 24 h 48 h 24 h 48 h

Control 8.24  0.01 a 8.18  0.02 a 0 0

Strawberry 8.21  0.03 a 8.13  0.02 a 0.4 0.6
Raspberry 8.14  0.03 b 8.02  0.03 b 1.2 2.0
Acai berry 7.94  0.03 c 7.81  0.03 c 3.6 4.5
Blueberry 7.86  0.03 d 7.69  0.05 d 4.6 6.0
Ocean Spray cranberry juice 7.30  0.01 e 7.20  0.02 e 11.4 11.9
Lakewood cranberry juice 7.17  0.03 f 6.97  0.02 f 12.9 14.8
CFU: colony forming unit and GAE: gallic acid equivalent.
Mean values followed by different letters within a column are significantly different (p < 0.05).

Table 4. The viable cell counts and percentage inhibition (compared to control) of S. aureus at two different bacterial
densities treated with pH-neutralized Lakewood cranberry juice at 86.6 mg GAE/mL bacterial culture for 24 h incubation
period

Staphylococcus aureus density Treatment Viable cell counts (log CFU/mL) Inhibition (%)

Undiluted (8.2 log CFU) Control 8.23  0.07 a 0

þCranberry 7.22  0.05 b 12.3
Diluted (5.0 log CFU) Control 5.00  0.05 c 0
þCranberry 2.83  0.09 d 43.4
CFU: colony forming unit and GAE: gallic acid equivalent.
a
Mean values followed by different letters are significantly different (p < 0.05).

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Food Science and Technology International 18(2)

juice was only slightly more potent in inhibiting bacterial
growth than cranberry juice, but both at least twice as
potent as blueberry. Acai berry was slightly less potent
than blueberry. Raspberry has only 10–13% of the
Lakewood cranberry antimicrobial activity, while
strawberry has no detectable activity (not significantly
different from control).
The increase in bacterial death as percentage inhib-
ition between the 24 and 48 h incubation periods for all
the berries were again small; thus, further studies were
concentrated on the 24 h incubation period.
Antimicrobial effects of cranberry at two
different densities of S. aureus
The maximum percentage inhibition of bacterial
growth that was achieved with the highest concentra-
tion of Ocean Spray and Lakewood cranberry juice
tested appears low, at 12.1% and 16.4% inhibition,
respectively (Tables 2 and 3). The bacterial density
could have affected the effectiveness of the cranberry
juice due to low phenolic content/cells ratio, as at
8.2 log CFU/mL, the bacterial density was twice the
level commonly used in similar studies in literature
(4.5 log CFU/mL). The high number of cells could
have also meant that the reproduction rate was much
faster than the cell death rate; therefore, the viable cell
counts did not decrease as much after being treated by
the cranberries. In order to determine whether the per-
centage inhibition could be increased, the bacterial
density was diluted by half. Indeed, lowering the cell
density from 8.2 log CFU/mL to 5.0 log CFU/mL to
increase the phenolic content/cells ratio resulted in
more than doubling of the percentage inhibition of
cell growth from 12.3% to 43.4% (Table 4).
However, the representation of results in percentage
inhibition could be misleading as a mere 12.3%
decrease from an initial bacterial concentration of
8.2 log CFU/mL would indicate the killing of 106
number of bacterial cells. This is a much larger
number of cell deaths compared to a 43.4% decrease
from an initial concentration of 5.0 log CFU/mL, which
equated to killing of 105 number of cells. This means
that while the antimicrobial effect of the cranberries
kills more cells at the higher density of 8.2 log CFU, it
is less sensitive in terms of percentage inhibition than
the lower density of 5.0 log CFU. But the conclusion
that cranberry is a potent antimicrobial against S.
aureus and the most potent among the berries studied
is inescapable.
Antimicrobial effects of solid-phase extracted
fractions of cranberry
In order to determine the nature of the antimicrobial
effect, the Lakewood cranberry juice was fractionated
into two fractions by solid-phase extraction using C18
solid phase. The column’s straight-through þ wash and
methanol-eluated fractions were obtained to determine
the association of antimicrobial activity with the
C18-bound materials, which is expected to contain the
phenolic materials. Indeed, the phenolic materials were
largely recovered in the methanol fraction, as indicated
by the total phenolic assay (Table 1), and were recov-
ered pH neutral and free of acids, salts, and sugars
which are not retained by the C18 solid phase.
The inhibitions of bacterial growth by the C18
retained materials obtained using 5.0 log CFU/mL of
bacterial density were 37.1%, 39.9%, and 42.5% for
43.3 86.6, and 173.2 mg GAE/mL of bacterial culture,
respectively (24 h incubation; Table 5). Indeed, the
percentage inhibition of the C18-bound materials was
similar to original juice at 39.9% versus 43.3%, respect-
ively (for the 86.6 mg GAE/mL bacterial culture tested;

Table 5. The viable cell counts and percentage inhibition (compared to control) of S. aureus treated with different vol-
umes of solid-phase extraction fractions of Lakewood cranberry juice for 24 h incubation period

Volume added to Phenolics concentration Viable cell counts

culture (mL) (mg GAE) (log CFU/mL) Inhibition (%)

Control 0 0 5.01  0.07 a 0

Straight-through fractiona 200 0 4.97  0.07 a ND
400 0 4.96  0.06 a ND
800 0 5.05  0.03 a ND
Methanol-eluated fraction 200 43.3 3.15  0.05 b 37.1
400 86.8 3.01  0.02 c 39.9
800 173.2 2.88  0.04 d 42.5
CFU: colony forming unit; GAE: gallic acid equivalent; and ND: none detected. Control, no C18 materials added.
Mean values followed by different letters are significantly different (p < 0.05).
a
The straight-through fraction contained no detectable phenolic materials; therefore, the values are displayed as 0 mg GAE.

184
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Tables 4 and 5). This provides strong evidence for the
antimicrobial activity residing within the C18-bound
phenolic materials. Doubling of phenolic materials
using the methanol fraction to 173.2 mgGAE/mL
increased the inhibition slightly to 42.5% (Table 5),
again indicating that near maximum inhibition was
attained.
In conclusion, the results from this study showed
that the Lakewood pure organic cranberry juice and
Ocean Spray cranberry cocktail juice possess potent
antimicrobial effect against the major food-borne
pathogen S. aureus. The Lakewood juice was slightly
more potent than the Ocean Spray juice at the same
amount of phenolic materials, and both cranberry
juices were much more potent than blueberry, acai
berry, and raspberry while strawberry has almost no
activity. The low pH of the berry juices and sugars,
acids, or salts were not responsible for the antimicrobial
effects, as indicated by the pH neutralization and solid-
phase extraction studies. Indeed, solid-phase extraction
showed that the antimicrobial activity was exclusively
associated with the C18-bound phenolic materials.
Given the rise of antibiotic resistant S. aureus and
other food-borne pathogenic bacteria, cranberry
shows promise as a natural and safe antimicrobial
for the food industry. Future challenges would include
identifying the nature of the phytochemical(s)
responsible for the observed antimicrobial activity
and elucidation of the antimicrobial mechanism(s).
FUNDING
This research is support by an internal DAFS funding.
ACKNOWLEDGMENT
The authors acknowledge Neslihan Goc’s contribution in
maintaining the bacterial cultures.
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