Letters in Applied Microbiology ISSN 0266-8254

ORIGINAL ARTICLE

Anti-biofilm activity of Quercus infectoria G. Olivier against

methicillin-resistant Staphylococcus aureus
S. Chusri1,2, P. Na Phatthalung2 and S.P. Voravuthikunchai2
1 Faculty of Traditional Thai Medicine, Prince of Songkla University, Hat Yai, Thailand
2 Natural Products Research Center and Department of Microbiology, Faculty of Science, Prince of Songkla University, Hat Yai, Thailand

Keywords Abstract
anti-biofilm activity, biofilm, cell surface
hydrophobicity, methicillin-resistant Aims: To establish the effect of Quercus infectoria G. Olivier extract and its
Staphylococcus aureus, Quercus infectoria. main constituent, tannic acid, on staphylococcal biofilm and their anti-biofilm
mechanisms.
Correspondence Methods and Results: Anti-biofilm activity of the plant materials on clinical
Supayang Piyawan Voravuthikunchai,
isolated of methicillin-resistant Staphylococcus aureus and methicillin-suscepti-
Department of Microbiology and Natural
Products Research Center, Faculty of Science,
ble Staph. aureus was employed using a crystal violet-stained microtiter plate
Prince of Songkla University, Hat Yai, method. The extract at minimum inhibitory concentration (MIC;
Songkhla 90112, Thailand. 0Æ25 mg ml)1) was significantly reduced the biofilm formation of the isolates
E-mail: [email protected] (P < 0Æ05). The effect on staphylococcal cell surface hydrophobicity (CSH) of
the test compounds was investigated as a possible mode of action of the anti-
2011 ⁄ 1177: received 14 July 2011, revised 9 biofilm activity. The hydrophobicity index of all the bacterial isolates increased
January 2012 and accepted 1 March 2012
following treatment with supra-MIC, MIC and sub-MIC of the extract and tan-
doi:10.1111/j.1472-765X.2012.03236.x
nic acid. Observation of the treated bacterial cells by electron microscopy
revealed that the test compounds caused clumps of partly divided cocci with
thickened and slightly rough cell wall.
Conclusions: The results indicated that Q. infectoria extract and tannic acid
affected staphylococcal biofilm formation and their effect on bacterial CSH and
cell wall may involve in the anti-biofilm activity.
Significance and Impact of the Study: This evidence highlighted the anti-
biofilm potency of the natural products and clarified their anti-biofilm
mechanisms.

2006). Many reports have proposed that the influence of

Introduction
anti-infective agents on CSH of bacterial cells would be
Methicillin-resistant Staphylococcus aureus (MRSA) is a important for anti-adhesion and anti-biofilm formation
harmful pathogen that is capable of causing diseases by of the treated pathogens as well (Fonseca et al. 2004;
utilizing a diverse armamentarium of virulent factors and Nithyanand et al. 2010). Decolonization agents such as
resistance mechanisms. Carriage of Staph. aureus is a risk chlorhexidine, mupirocin and triclosan have been used to
factor for subsequent infection and has been implicated eradicate nasal and hand carriage of methicillin-suscepti-
as a source of transmission (Butterly et al. 2010). The ble Staph. aureus (MSSA) and methicillin-resistant Staph-
adherence ability of micro-organisms to their host is ylococcus aureus (MRSA). Unfortunately, emergence of
mediated by a number of nonspecific factors (Lerebour resistant isolates as a result of long-term and intermittent
et al. 2004). Bacterial cell surface hydrophobicity (CSH) is usage of these decolonization agents has been frequently
one of the factors that govern initial bacterial adhesion to reported (Upton et al. 2003). Consequently, the discovery
various surfaces such as medical devices, teeth, contact of new anti-staphylococcal agents that could prevent
lenses and glass surface (Nostro et al. 2004; Prabu et al. adherence or biofilm formation remains an important

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Letters in Applied Microbiology 54, 511–517 ª 2012 The Society for Applied Microbiology 511
Q. infectoria: a potential anti-MRSA biofilm agent S. Chusri et al.

challenge to the scientific community and would be of

Biofilm formation
great use.
Extracts of Quercus infectoria G. Oliver nutgalls, a The isolates were evaluated for biofilm production accord-
traditional Thai medicinal plant with anti-ulceration ing to a modified microtiter plate method (Karaolis et al.
effect, have been investigated for anti-MRSA activity as 2005). An overnight culture grown in TSB (tryptic soy
well as antibacterial activity against skin pathogenic broth) supplemented with 0Æ25% glucose (GTSB) (200 ll)
bacteria (Chusri and Voravuthikunchai 2008; Meksee- was transferred to 96-well microtiter plate (Nunc, Wiesba-
pralard et al. 2010). Our previous investigation revealed den, Germany) and incubated at 37C for 24 h. The super-
that the ethanol extract of the nutgalls consists of tan- natant, including the planktonic cells, was removed, and
nins, flavonoids and steroidal compounds (Chusri and the wells were washed twice with phosphate-buffered saline
Voravuthikunchai 2009). The antibacterial activities as (PBS). The air-dried wells were stained with 0Æ1% crystal
well as the toxicity of the plant extract and tannic acid, violet (200 ll) for 30 min, and the plate was rinsed with
its main constituent with anti-MRSA activity are well PBS. The air-dried plates were added with 200 ll of
established (Umachigi et al. 2008; Chusri and Voravut- DMSO, and the OD at 570 nm was measured by microtit-
hikunchai 2009, 2011), while their effects on the er plate reader (Perkin-Elmer, Turku, Finland).
bacterial adhesion to host cells and biofilm production All strains were classified the cut-off OD (ODc) for the
are limited. Therefore, the aim of the investigation was microtiter plate test as three standard deviations above
to determine the ability of the extract from the nutgalls the mean OD of the negative control. The MRSA and
of Q. infectoria as well as tannic acid on the biofilm MSSA isolates were categorized as follows (Christensen
formation and the CSH of both MRSA from infective et al. 1985):
origins and MSSA from colonization origin to answer
whether any different effects occur between the bacteria
from different sources. OD £ ODc Nonadherent
ODc < OD £ 2ODc Weakly adherent
2ODc < OD £ 4ODc Moderately adherent
Materials and Methods 4ODc < OD Strongly adherent

Test bacterial isolates and anti-staphylococcal activity of

Quercus infectoria and its component
Effect of Quercus infectoria and tannic acid on the
Methicillin-resistant Staph. aureus, NPRC R001-R047, staphylococcal biofilm formation
were collected from patients at Hat Yai and Songklanaga-
rind hospitals, Hat Yai, Thailand. The bacterial isolates The biofilm formation of the representative MRSA and
were obtained from different sites of infectious including MSSA isolates after treatment with the ethanol extract
sputum, blood, pus and urine. Methicillin-susceptible Sta- and tannic acid was investigated by the microtiter plate
ph. aureus, NPRC S001-S050, were isolated from nasal method. Twofold serial dilutions of the extract and tannic
specimens of healthy volunteers at Department of Micro- acid were prepared in a 96-well microtiter plate in GTSB
biology, Faculty of Science, Prince of Songkla University, to obtain final concentrations of the test compounds
Hat Yai, Thailand. The reference strain, Staph. aureus ranged from MIC-1 ⁄ 8·MIC. One hundred and eighty
ATCC 25923, was used as a quality control. microliters of the bacterial suspension was transferred to
The powder of Q. infectoria nutgalls was prepared and 96-well microtiter plate. After incubation, the changes in
extracted with 95% ethanol as previously described biofilm-forming ability of treated cells were examined by
(Chusri and Voravuthikunchai 2009). The extract was the method as described earlier.
concentrated using vacuum rotary evaporator, completely
dried in an atmospheric oven and dissolved in 10% dim- Cell surface hydrophobicity (CSH)
ethylsulfoxide (DMSO; Merck, Darmstadt, Germany)
before use. Tannic acid that has been documented as the Cell hydrophobicity was measured by bacterial adherence
main constituent of the nutgalls (Kaur et al. 2008) and to hydrocarbon (BATH) according to the previous
possessed anti-MRSA activity (Chusri and Voravuthik- method (Nostro et al. 2004). Briefly, the bacterial suspen-
unchai 2009, 2011) was included as a positive control. A sion (100 ll), c. 5 · 106 CFU ml)1, was grown in 900 ll
modified microdilution method (CLSI 2009) was applied TSB at 37C for 18 h, and the cells were collected by cen-
to determine the minimum inhibitory concentration trifugation at 4000 g for 5 min. The pellets were washed
(MIC) of Q. infectoria extract and tannic acid against twice and then resuspended in 0Æ85% (w ⁄ v) sodium chlo-
both MRSA and MSSA isolates. ride to determine an initial optical density (OD initial) of

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512 Letters in Applied Microbiology 54, 511–517 ª 2012 The Society for Applied Microbiology
S. Chusri et al. Q. infectoria: a potential anti-MRSA biofilm agent

the cell suspension at 600 nm. The cell suspension (3 ml) Results
after the addition of toluene (0Æ25 ml) was agitated uni-
formly using a vortex mixer for 2 min and allowed to Anti-staphylococcal activity of Quercus infectoria extract
equilibrate at room temperature for 30 min. After toluene and tannic acid
phase was separated from the aqueous phase, the OD of
Antibacterial effects, expressed as MICs of the crude
the aqueous phase (OD final) was then determined at
extracts against MRSA, MSSA isolates and Staph. aureus
600 nm. The hydrophobicity index (HPBI) was calculated
ATCC 25923, were in the range of 0Æ13–0Æ50 mg ml)1
as: (OD initial-OD final ⁄ OD initial) · 100%. Low hydro-
(Table 1). Comparisons of the MIC values of the ethanol
phobic isolates were defined as a HPBI > 0 < 25, and
extract and tannic acid against MRSA and MSSA isolates
high hydrophobic isolates were classified as a HPBI ‡ 25
showed no difference.
(Akiyama et al. 1998).

Hydrophobic property and biofilm formation

Modification of staphylococcal CSH by the extract and
tannic acid The cell surface properties of both MRSA and MSSA iso-
lates are demonstrated in Table 2. The HPBI of MRSA
The properties of cell surface of the representative low
isolates was significantly lower than MSSA isolates
hydrophobic and high hydrophobic isolates after treat-
(P < 0Æ05). Most of MRSA isolates were classified as low
ment with the ethanol extract and tannic acid were inves-
hydrophobic strains, while 88% of MSSA isolates were
tigated by BATH test. The cell suspension (1Æ5 ·
described as high hydrophobic strains. A reference strain,
109 CFU ml)1) was incubated into 0Æ85% (w ⁄ v) NaCl
Staph. aureus ATCC 25923, was identified as a high
supplemented with the extract and tannic acid at 4·MIC,
hydrophobic strain.
2·MIC, MIC, 1 ⁄ 2·MIC, 1 ⁄ 4·MIC and 1 ⁄ 8·MIC at 37C
Table 3 presents the results of the biofilm formation of
for 24 h (10 : 1, v ⁄ v). The changes in cell surface hydro-
MRSA isolates from infective origins and MSSA isolates
phobicity of treated cells were observed by the method as
from carriers at 24 h. Most of the test isolates (>70%)
described earlier.

The effect of Quercus infectoria and its component on Table 2 Comparison of cell surface hydrophobicity of methicillin-
resistant Staphylococcus aureus (MRSA) and methicillin-susceptible
cell morphology of MRSA
Staph. aureus (MSSA) isolates
Methicillin-resistant Staphylococcus aureus cells were pre-
Percentage of tested isolates
treated with the extract and tannic acid at 4·MIC and
then prepared for the transmission electron microscope Hydrophobicity MRSA MSSA Staph. aureus
(TEM; JEM 100 CX II; JEOL,Tokyo, Japan) as described index (HPBI) (n = 47) (n = 50) ATCC 25923
previously (Chusri and Voravuthikunchai 2009). In addi- Low hydrophobicity 79 12
tion, glutaraldehyde-fixed cells were isolated on millipore (0 £ HPBI < 25)
filters and postfixed with 1% (w ⁄ v) osmium tetraoxide High hydrophobicity 21 88
at room temperature overnight. The bacterial cells were (25 £ HPBI £ 100)
dehydrated by passage through graded acetone ⁄ water Mean values ± SE 15Æ78 ± 14Æ04 55Æ26 ± 19Æ41* 81Æ44 ± 2Æ89
mixtures and treated with tetramethylsilane. The air- *Mean values are significantly different from MRSA isolates
dried cells were coated with gold and examined using (P < 0Æ05).
scanning electron microscopy (SEM; 5800LV, JEOL,
Tokyo, Japan).

Table 1 Minimum inhibitory concentrations (MICs) of Quercus infec- Table 3 Biofilm formation of methicillin-resistant Staphylococcus
toria ethanol extract and tannic acid against methicillin-resistant aureus (MRSA) and methicillin-susceptible Staph. aureus (MSSA) iso-
Staphylococcus aureus (MRSA) and methicillin-susceptible Staph. lates (modified microtiter plate method)
aureus (MSSA)
Adherence ability (% of the isolates)
MIC range ⁄ MIC90 (mg ml)1)
Test isolates* No Weak Moderate Strong
Antibacterial MRSA MSSA Staph. aureus
MRSA (n = 47) – 15 74 11
agents (n = 47) (n = 50) ATCC25923
MSSA (n = 50) – 18 78 4
Ethanol extract 0Æ13–0Æ50 ⁄ 0Æ25 0Æ13–0Æ50 ⁄ 0Æ25 0Æ13
Tannic acid 0Æ13–0Æ50 ⁄ 0Æ25 0Æ13–0Æ50 ⁄ 0Æ25 0Æ13 *Staphylococcus aureus ATCC 25923 was identified as strongly
adherent.

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Letters in Applied Microbiology 54, 511–517 ª 2012 The Society for Applied Microbiology 513
Q. infectoria: a potential anti-MRSA biofilm agent S. Chusri et al.

2·5 9
(a) (a)
8
2·0 * 7

OD 570 nm
* 6
OD 570 nm

1·5 * *
* * ** ** 5
*
1·0 4
* 3 *
* *
0·5 ** ** ** * ** 2 *
* * *
1
0·0 0
NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC
R015 R016 R029 R033 R034 R035 S010 S024 S028 S029 S042 S045
Methicillin resistant Staphylococcus aureus isolates Methicillin susceptible Staphylococcus aureus isolates

2·5 9
(b) 8 (b)
2·0 7

OD 570 nm
*
OD 570 nm

1·5 6
* 5
* *
1·0 * * * 4
* * *
* * * * * * 3
* *
0·5 * * * 2
1 * * *
0·0 0
NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC NPRC
R015 R016 R029 R033 R034 R035 S010 S024 S028 S029 S042 S045
Methicillin resistant Staphylococcus aureus isolates Methicillin susceptible Staphylococcus aureus isolates

Figure 1 The effect of Quercus infectoria ethanol extract (a) and tan- Figure 2 The effect of Quercus infectoria ethanol extract (a) and tan-
nic acid (b) at minimum inhibitory concentration (MIC, , 1 ⁄ 2·MIC nic acid (b) at minimum inhibitory concentration (MIC, ), 1 ⁄ 2·MIC
( ). 1 ⁄ 4·MIC ( ), 1 ⁄ 8·MIC ( ), and 1 ⁄ 16·MIC ( ) on the biofilm ( ), 1 ⁄ 4·MIC ( ), 1 ⁄ 8·MIC ( ), and 1 ⁄ 16·MIC ( ) on the biofilm
formation of representative methicillin-resistant Staphylococcus aureus formation of representative methicillin-susceptible Staphylococcus aur-
isolates, 1% DMSO ( ) was used as control. The means ± SDs for at eus isolates, 1% DMSO ( ) was used as control. The means ± SDs for
least duplicates are illustrated. *The biofilm formation of the isolates at least duplicates are illustrated. *The biofilm formation of the iso-
after treated with the compounds are significantly lower than the lates after treated with the compounds are significantly lower than
control (P < 0Æ05). the control (P < 0Æ05).

were classified as moderate adherence strains. About 10% MRSA isolates (Fig. 1b). Their effects on MSSA biofilm
of MRSA isolates were categorized as strong adherent, production are shown in Fig. 2. When exposed to sub-MIC
while only 4% of MSSA isolates were found with this of both the ethanol extract and tannic acid, there was no
ability. There was a negative correlation between the reduction in the biofilm formation of the MSSA isolates.
adherence ability and the cell surface properties of the test With the exception of MSSA NPRC S010, S024 and S028,
isolates (R2 = 0Æ093 and 0Æ001 for MRSA and MSSA, the inhibition of biofilm formation of the MSSA isolates
respectively). was observed after treatment with MIC of the ethanol
extract and tannic acid.
The effect of Quercus infectoria extract and tannic acid
on biofilm-forming ability of MRSA and MSSA The effect of Quercus infectoria extract and tannic acid
on MRSA and MSSA cell surface hydrophobicity
In the subsequent experiments, six representative MRSA
isolates (NPRC R015, R016, R029, R033, R034 and R035) In the following studies, the effect of supra-MICs, MIC
and six representative MSSA isolates (NPRC S010, S024, and sub-MICs of the plant extract and tannic acid on
S028, S029, S042 and S045) were selected as the most effi- CSH of representative high and low hydrophobic MRSA
cient isolates in terms of biofilm production. and MSSA isolates was investigated (Fig. 3). Supra-MIC
As presented in Fig. 1a, with the exception of MRSA values of both Q. infectoria extract and tannic acid signifi-
NPRC R015 and R033, all test concentrations (MIC- cantly increased the HPBI of low hydrophobic MRSA and
1 ⁄ 16·MIC) of the ethanol extract were significantly able to MSSA isolates (P < 0Æ05), while only low hydrophobic
reduce the biofilm formation of the test MRSA isolates MRSA isolates were significantly affected after treatment
(P < 0Æ05), while only at MIC and some sub-MICs of tan- with MIC of the ethanol extract and its constituent. Sub-
nic acid showed a decrease in biofilm formation of the test MICs values of these compounds demonstrated weak

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514 Letters in Applied Microbiology 54, 511–517 ª 2012 The Society for Applied Microbiology
S. Chusri et al. Q. infectoria: a potential anti-MRSA biofilm agent

100 100
90 (a) * * * 90 (b)
80 *
* * * * 80
70 * *
HPBI (%)
70

HPBI (%)
60 60 *
50 50 *
40 *
40
30 30
20 20
10 10
0 0
1% 4MIC 2MIC 1MIC 1/2 1/4 1/8 1% 4MIC 2MIC 1MIC 1/2 1/4 1/8
DMSO MIC MIC MIC DMSO MIC MIC MIC
Concentrations of ethanol extact and tannic acid Concentrations of ethanol extract and tannic acid

100 100
90 (c) 90 (d)
80 80
70
HPBI (%)

70

HPBI (%)
60 60
50 50
40 40
30 30
20 20
10 10
0 0
1% 4MIC 2MIC 1MIC 1/2 1/4 1/8 1% 4MIC 2MIC 1MIC 1/2 1/4 1/8
DMSO MIC MIC MIC DMSO MIC MIC MIC
Concentrations of ethanol extract and tannic acid Concentrations of ethanol extract and tannic acid

Figure 3 The effect of Quercus infectoria ethanol extract ( ) and tannic acid ( ) on the cell surface hydrophobicity of representative low hydro-
phobic methicillin-resistant Staphylococcus aureus (MRSA) isolate (a) and methicillin-susceptible Staph. aureus (MSSA) isolate (b) and high hydro-
phobic MRSA (c) and MSSA NPRC S019 (d), 1% DMSO (h) was used as control. *The hydrophobicity index of the isolates after treated with the
compounds are significantly different from the control (P < 0Æ05).

(a) (b) (c)

15 kV ×20,000 1 µm 180106 15 kV ×20,000 1 µm 180120 15 kV ×20,000 1 µm 210105

100 nm 100 nm 100 nm

Figure 4 Scanning and transmission electron micrographs showing the changes on cell surface of methicillin-resistant Staphylococcus aureus
grown in TSB containing the ethanol extract of Quercus infectoria (a) and tannic acid (b) at 4·MIC, for 12 h, 1% DMSO was used as control (c).

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Letters in Applied Microbiology 54, 511–517 ª 2012 The Society for Applied Microbiology 515
Q. infectoria: a potential anti-MRSA biofilm agent S. Chusri et al.

effect on the cell surface properties of these isolates. As O157: H7 cells treated with Q. infectoria exhibited a
demonstrated in Fig. 3c,d, the CSH of the high hydro- higher level of CSH than the surface of untreated cells as
phobic isolates was slightly altered following the treat- examined by BATH assay (Voravuthikunchai and Suwalak
ment of supra-MIC, MIC and sub-MIC values of both 2009). Tannic acid demonstrated similar results to the
the extract and tannic acid. plant extract, suggesting that it may be the composition
of the herb extract that influenced CSH. Fedtke et al.
(2007) have also demonstrated that Staph. aureus ypfP
The effect of Quercus infectoria extract and tannic acid
mutant with failure in lipoteichoic acid production dis-
on cell morphology of MRSA
played a significant increase in their HPBI but dramati-
Exposure of a representative MRSA to crude extract form cally lost their adhesion ability on polystyrene microtiter
Q. infectoria and tannic acid and observation under a plates.
SEM and a TEM revealed that the cells presented certain In conclusion, the effect of the extract on Staph. aureus
ultrastructural changes (Fig. 4a,b), in comparison with cell wall and cell surface hydrophobicity may cause a
the control in 1% DMSO which is shown in Fig. 4c. It reduction in staphylococcal biofilm formation. This scien-
was observed that the occurrence of these antibacterial tific evidence supports the traditional use of the plant,
agents caused slightly rough cell wall and increased the and in addition, the results of our study are helpful for
thickness of the cell wall when compared with the con- understanding the anti-biofilm mechanisms of these natu-
trol. ral products.

Discussion
Acknowledgements
Many reports have suggested that the influence of anti-
infective agents on CSH of bacterial cells would be impor- This work was supported by Agricultural Research
tant for anti-adhesion and anti-biofilm formation of the Development Agency (Public Organization) (fiscal year
treated organisms (Nostro et al. 2004; Prabu et al. 2006; 2011–2013) and the Thailand Research Fund: Royal
Walencka et al. 2007; Takahashi et al. 2010). Therefore, the Golden Jubilee, PhD Grant (PHD ⁄ 0006 ⁄ 2549, fiscal year
effects of the nutgall extract and tannic acid on biofilm for- 2006–2011).
mation and CSH have been investigated and discussed in
this report.
References
At the MIC value, Quercus infectoria extract significantly
reduced the biofilm formation of MRSA isolates (five of Akiyama, H., Yamasaki, O., Kanzaki, H., Tada, J. and Arata, J.
six isolates), while the reduction in the biofilm formation (1998) Adherence characteristics of Staphylococcus aureus
of the MSSA isolates was slightly lower (three of six iso- and coagulase-negative staphylococci isolated from various
lates). At sub-MIC, while the bacterial growth was not skin lesions. J Dermatol Sci 18, 132–136.
inhibited, the biofilm-forming ability of the organisms was Blanco, A.R., Sudano-Roccaro, A., Spoto, G.C., Nostro, A. and
significantly reduced. Other bioactive compounds such as Rusciano, D. (2005) Epigallocatechin gallate inhibits bio-
galloyl catechins and epicatechin gallate were reported to film formation by ocular staphylococcal isolates.
reduce slime production and inhibit the formation of Antimicrob Agents Chemother 49, 4339–4343.
Butterly, A., Schmidt, U. and Wiener-Kronish, J. (2010) Meth-
staphylococcal biofilm (Blanco et al. 2005). Even though
icillin-resistant Staphylococcus aureus colonization, its rela-
pseudomulticellular aggregates with thickened internal
tionship to nosocomial infection, and efficacy of control
bacterial cell wall were detected in MRSA treated with tan-
methods. Anesthesiology 113, 1453–1459.
nic acid, the compound was found to be inactive as an
Christensen, G.D., Simpson, W.A., Younger, J.J., Baddour,
anti-biofilm agent.
L.M., Barrett, F.F., Melton, D.M. and Beachey, E.H. (1985)
We found that the nature of CSH of infective and col- Adherence of coagulase-negative staphylococci to plastic
onization origin Staph. aureus was dramatically different; tissue culture plates: a quantitative model for the adher-
however, the plant extract increased the HPBI of all iso- ence of staphylococci to medical devices. J Clin Microbiol
lates. Some researchers have reported that sub-MIC values 22, 996–1006.
of certain antibiotics normally decrease the surface hydro- Chusri, S. and Voravuthikunchai, S.P. (2008) Quercus
phobicity and the ability of bacteria to adhere to epithe- infectoria: a candidate for the control of methicillin-
lial cells (Kustos et al. 2003). Contrastingly, studies on resistant Staphylococcus aureus infections. Phytother Res 22,
plant extracts revealed both enhancing and lowering of 560–562.
the CSH of bacteria (Dykes et al. 2003; Nostro et al. Chusri, S. and Voravuthikunchai, S.P. (2009) Detailed studies
2004; Prabu et al. 2006). The surface of Escherichia coli on Quercus infectoria Olivier (nutgalls) as an alternative

ª 2012 The Authors

516 Letters in Applied Microbiology 54, 511–517 ª 2012 The Society for Applied Microbiology
S. Chusri et al. Q. infectoria: a potential anti-MRSA biofilm agent

treatment for methicillin-resistant Staphylococcus aureus epidermidis to the Episkin reconstructed epidermis model
infections. J Appl Microbiol 106, 89–96. and to an inert 304 stainless steel substrate. J Appl Micro-
Chusri, S. and Voravuthikunchai, S.P. (2011) Damage of biol 97, 7–16.
staphylococcal cytoplasmic membrane by Quercus infectoria Mekseepralard, C., Kamkaen, N. and Wilkinson, J.M. (2010)
G. Olivier and its components. Lett Appl Microbiol 52, Antimicrobial and antioxidant activities of traditional Thai
565–572. herbal remedies for aphthous ulcers. Phytother Res 24,
CLSI (2009) M07-A8-Methods for Dilution Antimicrobial 1514–1519.
Susceptibility Tests for Bacteria That Grow Aerobically; Nithyanand, P., Thenmozhi, R., Rathna, J. and Pandian, S.K.
Approved Standard-Eighth Edition. Wayne, PA, USA: Clini- (2010) Inhibition of Streptococcus pyogenes biofilm forma-
cal and Laboratory Standards Institute. tion by coral-associated actinomycetes. Curr Microbiol 60,
Dykes, G.A., Amarowicz, R. and Pegg, R.B. (2003) An antioxi- 454–460.
dant bearberry (Arctostaphylos uva-ursi) extract modulates Nostro, A., Cannatelli, M.A., Crisafi, G., Musolino, A.D.,
surface hydrophobicity of a wide range of food-related Procopio, F. and Alonzo, V. (2004) Modifications of
bacteria: implications for functional food safety. Food hydrophobicity, in vitro adherence and cellular aggregation
Control 14, 515–518. of Streptococcus mutans by Helichrysum italicum extract.
Fedtke, I., Mader, D., Kohler, T., Moll, H., Nicholson, G., Lett Appl Microbiol 38, 423–427.
Biswas, R., Henseler, K., Gotz, F. et al. (2007) A Staphylococ- Prabu, G.R., Gnanamani, A. and Sadulla, S. (2006) Guaijaverin
cus aureus ypfP mutant with strongly reduced lipoteichoic -a plant flavonoid as potential antiplaque agent against
acid (LTA) content: LTA governs bacterial surface proper- Streptococcus mutans. J Appl Microbiol 101, 487–495.
ties and autolysin activity. Mol Microbiol 65, 1078–1091. Takahashi, H., Suda, T., Tanaka, Y. and Kimura, B. (2010)
Fonseca, A.P., Extremina, C., Fonseca, A.F. and Sousa, J.C. Cellular hydrophobicity of Listeria monocytogenes involves
(2004) Effect of subinhibitory concentration of piperacil- initial attachment and biofilm formation on the surface of
lin ⁄ tazobactam on Pseudomonas aeruginosa. J Med Micro- polyvinyl chloride. Lett Appl Microbiol 50, 618–625.
biol 53, 903–910. Umachigi, S.P., Jayaveera, K.N., Ashok Kumar, C.K., Kumar,
Karaolis, D.K., Rashid, M.H., Chythanya, R., Luo, W., Hyodo, G.S., Vrushabendra-swamy, B.M. and Kishore Kumar,
M. and Hayakawa, Y. (2005) c-di-GMP (3’-5’-cyclic digu- D.V. (2008) Studies on wound healing properties of
anylic acid) inhibits Staphylococcus aureus cell-cell interac- Quercus infectoria. Trop J Pharm Res 7, 913–919.
tions and biofilm formation. Antimicrob Agents Chemother Upton, A., Lang, S. and Heffernan, H. (2003) Mupirocin
49, 1029–1038. and Staphylococcus aureus: a recent paradigm of emerging
Kaur, G., Athar, M. and Alam, M.S. (2008) Quercus infectoria antibiotic resistance. J Antimicrob Chemother 51,
galls possess antioxidant activity and abrogates oxidative 613–617.
stress-induced functional alterations in murine macrophag- Voravuthikunchai, S.P. and Suwalak, S. (2009) Changes in cell
es. Chem Biol Interact 171, 272–282. surface properties of shiga toxigenic Escherichia coli by
Kustos, T., Kustos, I., Kilar, F., Rappai, G. and Kocsis, B. Quercus infectoria G. Olivier. J Food Prot 72, 1699–1704.
(2003) Effect of antibiotics on cell surface hydrophobicity Walencka, E., Rozalska, S., Wysokinska, H., Rozalski, M.,
of bacteria causing orthopedic wound infections. Chemo- Kuzma, L. and Rozalska, B. (2007) Salvipisone and
therapy 49, 237–242. aethiopinone from Salvia sclarea hairy roots modulate
Lerebour, G., Cupferman, S. and Bellon-Fontaine, M.N. (2004) staphylococcal antibiotic resistance and express anti-
Adhesion of Staphylococcus aureus and Staphylococcus biofilm activity. Planta Med 73, 545–551.

ª 2012 The Authors

Letters in Applied Microbiology 54, 511–517 ª 2012 The Society for Applied Microbiology 517