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

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 from 100% fruit juice with no added water or sugar)
in plants against free radicals and radiation damage and acai berry, blueberry, strawberry, and raspberry
(Stevenson and Hurst, 2007). Dark-colored berry such fruits were obtained from a local supermarket. Sep-
as cranberry represents one of the most important diet- Pak C18 cartridges were from Waters Corporation.
ary sources of phenolic compounds that can be categor- Folin–Ciocalteu ReagentÕ (2N) and gallic acid were
ized into four main groups: simple phenolic acids, such from Sigma–Aldrich Inc. (St. Louis, MO, USA). All
as gallic acid and caffeic acid, lignin fragments and other common laboratory chemicals used were of ana-
monomeric flavonoids (catechins, flavonols, and antho- lytical grade or better. Water used was MilliporeÕ deio-
cyanins) and polymeric (proanthocyanidins; PCA) nized water.
flavonoids (Fang et al., 2009; Singh et al., 2009).
The American cranberry fruit, Vaccinium macrocar- Bacterial culture. Staphylococcus aureus strain
pon, is a traditional fruit originating from America and ACTT25923 (Seattle) was kept under refrigeration at
was historically used by Native Americans as a health 4  C as stock culture and reactivated weekly to main-
promoting fruit (Apostolidis et al., 2008). Cranberry tain viability. To reactivate the culture, a single isolated
has been shown to possess potent antioxidant activity colony was obtained from the stock culture plate and
and antimicrobial properties linked to its phenolic inoculated into 9 mL of brain heart infusion (BHI)
contents (Guo et al., 2007; Lacombe et al., 2010; broth and incubated at 37  C for 24 h. An inoculation
Nohynek et al., 2006). Cranberry, like most other loop is then used to prepare a streak plate from the
berries, contains B-type PAC, which is a short-chain incubated BHI and the plate incubated for another
polymer of anthocyanidin units containing multiples 24 h at 37  C before refrigeration as further stock
of the basic structure linked by a single ether bond culture.
(Dixon et al., 2005). However, cranberry also contains
a unique A-type PAC which bears a second ether Culture media. Baird Parker agar (BPA) plates were
linkage through the chroman’s oxygen (Howell, 2007). prepared for the purpose of enumeration of bacteria.
A-type PAC is linked to antimicrobial effect in prevent- BPA was prepared following the protocol specified by
ing urinary tract infection by P-fimbriated Escherichia the manufacturer’s instructions. The media powder
coli (Howell, 2007; Johnson-White et al., 2006; Liu used was purchased from Oxoid. The required
et al., 2006, 2008). The inhibition mechanism appears amount of powdered media was added to 1 L of dis-
to be due to the ability of A-type PAC molecules tilled water in Schott bottles and boiled to insure that
inhibiting the attachment of E. coli to the epithelial the powder was well mixed and dissolved. The bottles
cells lining of the urinary tract, preventing infection. were then autoclaved at 121  C for 15 min and allowed
Like all bacterial infections, antibiotic resistance is a to cool to 50  C before adding 50 mL of egg yolk tel-
major health threat and naturally occurring antibacter- lurite emulsion. The bottle was gently swirled to insure
ial agents are highly sort after (Foxman et al., 2000). even distribution and then aseptically poured into ster-
Recently, cranberry extracts have also been shown to ile Petri dishes. The plates were then refrigerated at 4  C
display antimicrobial effects on S. aureus, Listeria until further use.
monocytogenes, and E. coli strain by as yet undermined BHI broth was also prepared by evenly mixing the
mechanism of killing bacteria or inhibiting growth powdered broth purchased from Oxoid. The required
(Lacombe et al., 2010; Wu et al., 2009). amount of powder to make up 1 L of broth was 37 g.
This article compares the antimicrobial effect of two The mixture was well shaken to insure that the powder
commercially available cranberry juice products against had dissolved and mixed evenly. The broth (9 mL) was
S. aureus and compares the cranberry juices with the then distributed into individual McCartney bottles with
extracts of acai berry, blueberry, raspberry, and straw- a Zipette Classic bottle-top dispenser. The bottles were
berry fruits. We also provided evidence for the anti- placed in racks and autoclaved at 121  C for 15 min and
microbial effect of cranberry residing with its phenolic allowed to cool. The bottles were refrigerated at 4  C
materials. until further use.
Peptone water at a concentration of 0.01% w/v was
MATERIALS AND METHODS prepared for the purpose of dilution. The powdered
media was also obtained from Oxoid. One gram of
Materials
powder was evenly dissolved in 1 L of distilled water.
Ocean SprayÕ cranberry juice cocktail light (manufac- The peptone water (9 mL) was then distributed into
turer stated ingredients: water, 27% cranberry juice (v/v individual McCartney bottles. The bottles were placed
from concentrates)), natural flavors (not specified), arti- in racks and autoclaved at 121  C for 15 min and
ficial sweetener (sucralose)), LakewoodÕ pure organic allowed to cool. The bottles were refrigerated at 4  C
cranberry juice (manufacturer stated ingredient: made until further use.

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Lian et al.

Sterilized berry samples. Acai berry, blueberry, rasp- mean absorbance (AU) versus mass (mg) from the gallic
berry, and strawberry juices were prepared from whole acid standard plot: AU ¼ 0.0088 mg (sample) þ 0.0146
fruits by blending individual berry with a commercial (R2 ¼ 0.9973).
blender and centrifugation in a bench-top centrifuge at
5000 r/min for 60 min to obtain clear juice. Antimicrobial effects of berry extracts. Staphylococcus
The original pH values of the various berry samples aureus was cultured by taking an isolated colony from a
were fairly acidic; Lakewood cranberry (1.19), Ocean refrigerated stock plate and inoculating it into 9 mL of
Spray cranberry (2.96), acai berry (4.32), blueberry BHI. The broth was incubated at 37  C for 24 h. To
(3.34), strawberry (3.69), and raspberry (3.23). obtain the initial bacterial count, 1 mL of the cultured
Lakewood cranberry has the strongest acidity and BHI was used to conduct 6  1 : 10 serial dilutions using
required 0.5 M of NaOH (final concentration in peptone water. Duplicate 100 mL of the 106 diluted
juice) to neutralize. Ocean Spray cranberry required sample was then plated on BPA using the spread
0.15 M while the other berries required less than plate method. The duplicate plates were incubated at
0.1 M of NaOH to neutralize. 37  C for 24 h and their bacterial colonies counted.
Original (acidic) and pH neutralized (pH 7) juices For the antimicrobial effect study, 1 mL of the bac-
were individually sterilized by passage through sterile terial culture in BHI was pipetted out into separate
Acrodisc 25 mm filter with a 0.2 mm Supor membrane. 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
Methods
sample was taken from each bottle and subjected to
Partially purified cranberry. A sample of the Lakewood 5  1 : 10 serial dilutions with peptone water.
cranberry juice was subjected to solid-phase extraction Duplicate 100 mL was taken from the 103, 104, and
using a 30 mL Sep-Pak C18 solid-phase column. The 105 diluted samples and were plated on separate BPA
juice (50 mL) was applied to the column and washed plates. The plates were incubated at 37  C for 24 h and
with 10 volume of water before elution with 5 bacterial colonies counted to obtain the 24 h antimicro-
volume of 100% methanol to recover the intensely col- bial effect. For 48 h antimicrobial effect, the bottles
ored bound materials. The straight through and water were further incubated for another 24 h and duplicate
wash were combined, pH adjusted to neutral with 100 mL samples were again taken from each bottle for
NaOH, solution evaporated to dryness at 40  C under plating and incubation on BPA for 24 h and colonies
reduced pressure, and the dried materials reconstituted counted.
with 25 mL water (2 concentrated). The methanol
eluate was similarly dried without pH adjustment (solu- Statistical analysis. Each determination was performed
tion was neutral) and materials also reconstituted with in duplicate. The number of bacterial cells were
25 mL water (2  concentrated). Both samples were reported as log CFU/mL (CFU, colony forming unit)
sterilized by passage through 0.2 mm membrane filter and treatments were assigned for comparison. Analyses
and stored at 4  C before analysis. of variance were performed on the mean and range of
the results using the SAS General Linear Model pro-
Quantification of phenolic contents. Samples (berries cedures with SAS 9.2 software (Statistical Analysis
juices and cranberry C18 straight through and metha- System Institute, Inc., Cary, NC, USA). Differences
nol eluates) were analyzed for their total phenolic con- among treatments were examined for the level of sig-
tents using Folin–Ciocalteu Reagent and gallic acid as nificance by least significant difference. Significance of
standards as described by Slinkard and Singleton differences was defined as p < 0.05.
(1977). One milliliter of sample or standard solution
was mixed with 1 mL of freshly diluted Folin– RESULTS AND DISCUSSION
Ciocalteu Reagent (1 : 10 dilutions with water; 0.2 N)
Phenolic contents of berry samples
in a plastic tube and shaken vigorously. After 10 min,
3 mL of Na2CO3 solution (10%, w/v) was added and Table 1 presents the phenolic contents of the berry sam-
the mixture incubated at 40  C for 15 min with intermit- ples as determined by a colorimetric method using
tent shaking. The absorbance was measured using a Folin–Ciocalteu Reagent and expressed as GAE
UV/vis spectrophotometer (UV-1601 Shimadzu) at (Slinkard and Singleton, 1977). It can be seen that the
765 nm. A range of varying amounts of the sample Lakewood organic 100% cranberry concentrate con-
(n ¼ 4) or gallic acid standard (n ¼ 5) were determined. tained more than twice the amount of phenolic materials
The concentration of phenolic materials as gallic acid than Ocean Spray cranberry light cocktail. This is due to
equivalent (GAE) was calculated according to the fol- the Ocean Spray cranberry containing only 27% of juice.
lowing linear equation that was obtained from a plot of 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 Not surprisingly, all the phenolic materials were recov-
phenolic materials. The phenolic contents were not ered in the C18-bound methanol-eluated fraction
related to the weight of fruits as we are only concerned (Table 1) as they have affinity for the C18 solid phase.
with relating the phenolic contents (as GAE) to their
antimicrobial effects on the viability of S. aureus.
Antimicrobial effects of Ocean Spray and
A sample of the Lakewood cranberry concentrate
Lakewood cranberry juice on S. aureus
was fractionated by solid-phase extraction using a
C18 solid phase open column into straight-through The antimicrobial effects of Ocean Spray and
and methanol-eluated fractions. The fractions were Lakewood cranberry juice were tested against S.
reconstituted to twice their original concentrations aureus. Five different concentrations (in terms the phen-
(i.e., half the original volume applied to the column). 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-
Table 1. The total phenolic content of juices made with
berries
iods. The results (Table 2) showed that the percentage
inhibition of S. aureus growth, as measured by viable
Berry samples mg GAE/mL cell counts, compared to control without added juice
was greater in Lakewood than the Ocean Spray cran-
Ocean Spray cranberry juice 867  1
berry juice at the same concentration of phenolic mater-
Lakewood cranberry juice 2305  11
ials for a concentration range 0–86.6 mg GAE/mL
Acai berry 1006  4 bacterial culture tested. Higher concentration
Strawberry 743  9 Lakewood cranberry juice at 173.1 mg GAE/mL only
Blueberry 2018  8 increases the percentage inhibition slightly, indicating
Raspberry 771  1 optimal concentration effect has been reached (the
Lakewood cranberry C18 ND Ocean Spray cranberry juice could only be added to
straight-through fractiona 86.6 mg GAE/mL in the assay protocol).
Lakewood cranberry C18 4585  15 The inhibition percentage, or number of cell deaths,
methanol-eluated fractiona only increased slightly between the 24 and 48 h incuba-
GAE: gallic acid equivalent and ND: none detected. tion period for all concentrations (Table 2), indicating
a
Extraction fractions reconstituted in half the volume of original that a 24 h incubation period would be sufficient for
sample applied to C18 column. 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).

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Lian et al.

culture, the inhibition achieved was 12.3–14.9% for further supported this inference as the differences in
Lakewood and 11.5–13.2% for Ocean Spray cranberry the overall inhibition of the pH-adjusted cranberry
juice (Table 2). Both juice samples contain some ori- treatments as compared to their original pH was only
ginal sugars attributed to the fruit source that might less than 1.5%. The high acidity of cranberry juice is
contribute to promoting some degree of bacterial believed to be due to the presence of organic acids (Van
growth. However, the flavoring and pectin additives Immerseel et al., 2006). In the case of the Lakewood
in the Ocean Spray cranberry juice might have compli- cranberry juice, the pH of the juice was very low, 1.19.
cated further the antimicrobial effect by promoting
some bacterial growth that produces a lower percentage
Antimicrobial effects of various berries on
inhibition observed, compared to the Lakewood cran-
S. aureus
berry juice which was made with 100% pure fruit juice
with no additives added. In order to establish the effectiveness of cranberry as an
Importantly, the results also showed that the anti- antimicrobial, an inhibition experiment was carried out
microbial effects were not due to the strong acidity of to compare both Lakewood and Ocean Spray cranberry
the cranberry juices as the percentage inhibitions were juices against four other berry juices prepared from
similar for original and pH-neutralized juices across all whole fruits (acai berry, blueberry, raspberry, and straw-
concentrations tested (Table 2). Early studies have berry). The berry juices were tested at their neutralized
credited the antimicrobial effects of cranberry extracts pH (7) and were administered at 86.6 mgGAE/mL bac-
to the extreme acidity of the juice (Johnson-White terial culture for 24 and 48 h period incubation period.
et al., 2006). However, it has since been demonstrated The results (Table 3) showed that the order of antimicro-
by a couple of later studies (Lacombe et al., 2010; Wu bial activity against S aureus was Lakewood cranberry
et al., 2008) that pH does not appear to play a central juice > Ocean Spray cranberry juice  blueberry > acai
role in the antimicrobial effects of berries. Our data 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 decrease from an initial bacterial concentration of
growth than cranberry juice, but both at least twice as 8.2 log CFU/mL would indicate the killing of 106
potent as blueberry. Acai berry was slightly less potent number of bacterial cells. This is a much larger
than blueberry. Raspberry has only 10–13% of the number of cell deaths compared to a 43.4% decrease
Lakewood cranberry antimicrobial activity, while from an initial concentration of 5.0 log CFU/mL, which
strawberry has no detectable activity (not significantly equated to killing of 105 number of cells. This means
different from control). that while the antimicrobial effect of the cranberries
The increase in bacterial death as percentage inhib- kills more cells at the higher density of 8.2 log CFU, it
ition between the 24 and 48 h incubation periods for all is less sensitive in terms of percentage inhibition than
the berries were again small; thus, further studies were the lower density of 5.0 log CFU. But the conclusion
concentrated on the 24 h incubation period. that cranberry is a potent antimicrobial against S.
aureus and the most potent among the berries studied
is inescapable.
Antimicrobial effects of cranberry at two
different densities of S. aureus
Antimicrobial effects of solid-phase extracted
The maximum percentage inhibition of bacterial
fractions of cranberry
growth that was achieved with the highest concentra-
tion of Ocean Spray and Lakewood cranberry juice In order to determine the nature of the antimicrobial
tested appears low, at 12.1% and 16.4% inhibition, effect, the Lakewood cranberry juice was fractionated
respectively (Tables 2 and 3). The bacterial density into two fractions by solid-phase extraction using C18
could have affected the effectiveness of the cranberry solid phase. The column’s straight-through þ wash and
juice due to low phenolic content/cells ratio, as at methanol-eluated fractions were obtained to determine
8.2 log CFU/mL, the bacterial density was twice the the association of antimicrobial activity with the
level commonly used in similar studies in literature C18-bound materials, which is expected to contain the
(4.5 log CFU/mL). The high number of cells could phenolic materials. Indeed, the phenolic materials were
have also meant that the reproduction rate was much largely recovered in the methanol fraction, as indicated
faster than the cell death rate; therefore, the viable cell by the total phenolic assay (Table 1), and were recov-
counts did not decrease as much after being treated by ered pH neutral and free of acids, salts, and sugars
the cranberries. In order to determine whether the per- which are not retained by the C18 solid phase.
centage inhibition could be increased, the bacterial The inhibitions of bacterial growth by the C18
density was diluted by half. Indeed, lowering the cell retained materials obtained using 5.0 log CFU/mL of
density from 8.2 log CFU/mL to 5.0 log CFU/mL to bacterial density were 37.1%, 39.9%, and 42.5% for
increase the phenolic content/cells ratio resulted in 43.3 86.6, and 173.2 mg GAE/mL of bacterial culture,
more than doubling of the percentage inhibition of respectively (24 h incubation; Table 5). Indeed, the
cell growth from 12.3% to 43.4% (Table 4). percentage inhibition of the C18-bound materials was
However, the representation of results in percentage similar to original juice at 39.9% versus 43.3%, respect-
inhibition could be misleading as a mere 12.3% 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.

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Lian et al.

Tables 4 and 5). This provides strong evidence for the Dixon RA, Xie DY and Sharma SB. (2005).
antimicrobial activity residing within the C18-bound Proanthocyanidins – a final frontier in flavonoid research?
phenolic materials. Doubling of phenolic materials New Phytologist 165(1): 9–28.
using the methanol fraction to 173.2 mgGAE/mL Fang Z, Zhang Y, Lu Y, Ma G, Chen J and Liu DXY. (2009).
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attained. (2000). Urinary tract infection: self-reported incidence and
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Ocean Spray cranberry cocktail juice possess potent et al. (2007). Iron-binding properties of plant phenolics
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amount of phenolic materials, and both cranberry their role in prevention of urinary tract infections.
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Johnson-White B, Buquo L, Zeinali M and Ligler S. (2006).
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Prevention of nonspecific bacteria cell adhesion in
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showed that the antimicrobial activity was exclusively chemicals of selected clonal herbs species of Lamiaceae
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shows promise as a natural and safe antimicrobial Antimicrobial action of the American cranberry constitu-
for the food industry. Future challenges would include ents; phenolics, anthocyanins, and organic acids, against
E. coli O157:H7. International Journal of Food
identifying the nature of the phytochemical(s)
Microbiology 139(1–2): 102–107.
responsible for the observed antimicrobial activity
Liu Y, Black MA, Caron L and Camesano T. (2006). A role
and elucidation of the antimicrobial mechanism(s). of cranberry juice on molecular-scale surface characteris-
tics and adhesion behavior of Escherichia coli.
FUNDING Biotechnology and Bioengineering 93(2): 297–305.
Liu Y, Gallardo-Moreno M, Pinzon-Arango PA, Reynolds
This research is support by an internal DAFS funding.
Y, Rodriguez G and Camesano TA. (2008). Cranberry
changes the physicochemical surface properties of E. coli
ACKNOWLEDGMENT and adhesion with uroepithelial cells. Colloids and Surfaces
The authors acknowledge Neslihan Goc’s contribution in B: Biointerfaces 65(1): 35–42.
maintaining the bacterial cultures. Nohynek LJ, Alakomi H, Kahkonen MP, Heinonen M,
Helander IM, Oksman-Caldentey K, et al. (2006).
Berries phenolics: antimicrobial properties and mechan-
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