Letters in Applied Microbiology 2005, 41, 476–481 doi:10.1111/j.1472-765X.2005.01790.

Triclosan–bacteria interactions: single or multiple target sites?

M. Gomez Escalada1, A.D. Russell1, J.-Y. Maillard1 and D. Ochs2

1
Welsh School of Pharmacy, Cardiff University, Cardiff, UK and 2Ciba Specialty Chemicals Inc., Grenzach-Wyhlen, Germany

2005/0010: received 5 January 2005, revised 10 May 2005 and accepted 30 June 2005

ABSTRACT
M . G O M E Z E S C A L A D A , A . D . R U S S E L L , J . - Y . M A I L L A R D A N D D . O C H S . 2005.
Aims: To investigate the inhibitory and lethal effects of triclosan against several micro-organisms at different stages
of their phase of population growth.
Methods and Results: Triclosan minimum inhibitory concentrations against several test organisms were
determined in broth and agar using standard protocols. The bisphenol effect on bacterial population growth kinetics
was studied using the Bioscreen C microbial growth analyser. Finally, the efficacy of triclosan on phases of bacterial
growth was determined using a standard suspension test. The duration of the lag phase for all micro-organisms
tested was increased by bisphenol in a concentration-dependent manner. The population growth kinetics of the
micro-organisms was also altered after biocide exposure. At higher concentrations, triclosan was bactericidal
regardless of their phase of population growth, although population in stationary phase and particularly, washed
suspensions, were more resilient to the lethality of triclosan. This lethal activity was concentration and contact time
dependent, and in some instances, bactericidal activity of bisphenol was observed within 15 s.
Conclusions: Low concentrations of triclosan affected the growth of several bacteria, while higher concentrations
were bactericidal regardless of the bacterial phase of population growth.
Significance and Impact of the Study: Here, we presented clear evidence that the interaction of triclosan with
the bacterial cell is complex and its lethality cannot be explained solely by the inhibition of metabolic pathways such
as the enoyl acyl-reductase. However, the inhibition of such pathways cannot be ruled out as part of the lethal
mechanism of the bisphenol at a low bactericidal concentration.

Keywords: antibacterial activity, concentration, growth phase, triclosan

micro-organisms such as methicillin-resistant Staphylococ-

INTRODUCTION
Triclosan (Irgasan DP 300; 2,4,4¢ trichloro-2¢-hydroxydi-
phenyl ether) is a chlorinated diphenyl ether or bisphenol
that possesses high antibacterial activity particularly
against many Gram-positive and negative bacteria
(McDonnell and Russell 1999). As a result of its broad-
spectrum antimicrobial properties and its safety profile
(Bhargava and Leonard 1996), triclosan is widely used in
personal health-care products (Jones et al. 2000) and has
been effectively used clinically to eradicate troublesome
Correspondence to: J.-Y. Maillard, Welsh School of Pharmacy, Cardiff University,
Cardiff CF10 3XF, UK (e-mail: maillardj@cardiff.ac.uk).
cus aureus (Brady et al. 1990; Webster et al. 1994; Zafar
et al. 1995).
Triclosan was found to target fatty acid synthesis by
inhibiting the enzyme enoyl reductase (enoyl-acyl carrier
protein reductase, FabI) (McMurry et al. 1998a). It acts as a
potent irreversible inhibitor by mimicking its natural
substrate (Heath et al. 1998; Levy et al. 1999) and this
inhibition has been described as being slow and competitive
(Heath et al. 1999). At higher concentration, the bisphenol
is likely to damage the bacterial membrane (Villalain et al.
2001). Although the antimicrobial activity of the bisphenol
and most likely its interaction(s) with the cell depends
ª 2005 The Society for Applied Microbiology

heavily upon its concentration (Russell 2004), it remains
important to substantiate its bactericidal activity regardless
of the cell metabolic status.
Here, the activity of triclosan at low and high concentra-
tions was investigated against several bacterial genera at
different phases of population growth, in order to determine
the importance of bacterial metabolism for its bacteriostatic
and bactericidal activity.
MATERIALS AND METHODS
Test organisms
The organisms used in this investigation were those
recommended by the European Standard for Chemical
Disinfectants and Antiseptics (CEN 1997a): Escherichia coli
(ATCC 10536), Enterococcus hirae (ATCC 8191) and
Staphylococcus aureus (ATCC 6538). However, Pseudomonas
aeruginosa was not selected because it is intrinsically resistant
to triclosan. All strains were maintained on tryptone soya
agar (TSA; Oxoid, Basingstoke, UK) and grown overnight
in 10 ml tryptone soya broth (TSB; Oxoid) at 37
C.
Bacterial stock cultures were kept at 4
C and renewed every
fortnight from a frozen culture kept at )20
C in 50%
glycerol (Fisher, Loughborough, UK).
Viable bacteria were enumerated using an adaptation from
the Miles–Misra method. Briefly, 100 ll of the bacterial
sample was serially diluted in 900 ll phosphate buffered
solution (PBS; Sigma, Poole, UK) and three 10 ll drops of
each dilution were spotted on the surface of a TSA plate.
After incubation at 37
C for 24 h, spots that contained
between 3 and 60 colonies were counted and the bacterial
concentration was expressed as CFU ml)1.
Minimum inhibitory concentration in broth
and agar
Triclosan was obtained from Ciba Specialty Chemicals
(Grenzach-Wyhlen, Germany). Stock solutions of triclosan
of 10 000, 1000, 100 and 10 mg l)1 were prepared using
dimethylsulfoxide (DMSO; Sigma) as a solvent. Further
dilutions were made in sterile distilled water or culture
medium, as appropriate.
For the determination of broth minimum inhibitory
concentrations (MICs), a series of triclosan concentrations
prepared in TSB were inoculated with 10 ll (approx.
1–5 · 104 CFU) of an overnight culture of the test organ-
ism. For agar MICs, a series of concentrations of triclosan
prepared in TSA was inoculated with a 1 ll drop (approx.
1–5 · 105 CFU) of an overnight culture of the test organism
using the Denley Multipoint Inoculator (Denley, Billing-
hurst, UK). After incubation of bottles or plates at 37
C for
24 h, the MIC was taken as the lowest concentration of
ª 2005 The Society for Applied Microbiology, Letters in Applied Microbiology, 41, 476–481, doi:10.1111/j.1472-765X.2005.01790.x
TRICLOSAN INTERACTIONS WITH BACTERIA 477
triclosan that inhibited visible growth. The final concentra-
tion of DMSO did not exceed 1% and had no growth-
inhibitory properties on any cultures tested.
Inhibitory effects on phases of population growth
The effect of subinhibitory concentrations of triclosan on
the test organisms was determined spectrophotometrically
using the Bioscreen C analyser (Labsystems, Helsinki,
Finland). A 150-ll aliquot of a range of subinhibitory
concentrations of triclosan was added to the Bioscreen plate
(Labsystems) with 150 ll (approx. 1–5 · 105 CFU) of an
overnight culture of the test organism. Appropriate solvent
and negative controls were also included. The plates were
incubated at 37
C and optical density (OD) readings taken
at 540 nm every 10 min for 24 h.
Lethal effect on washed cells and on cells
at different phases of population growth
The lethal effect of triclosan was determined as follows:
overnight cultures of the test organism were washed three
times by centrifugation (1500 · g for 15 min) with resus-
pension in sterile distilled water. A 1-ml sample of each
washed suspension was added to a range of triclosan
concentrations and after 15 s, 2, 5, 10, 15 and 20 min, a
1-ml aliquot was placed in 9 ml of neutralizing solution.
Samples were left in neutralizer for at least 5 min, serially
diluted and a viable count performed as described above.
In order to be consistent with the inoculum size used with
the washed suspension, a standard curve was produced (data
not shown). Briefly, 5 ml of a washed suspension of bacteria
resuspended in water was added to 5 ml of double-strength
TSB and incubated in a shaking water bath at 37
C. At
5-min intervals for 30 min, a 100 ll sample was removed
and the viability was assessed using the drop counting
method. To maintain the same inoculum and in order for
the cells to reach their exponential growth phase, it was
found that E. coli, Ent. hirae and S. aureus had to be
incubated for 12Æ5, 17Æ5 and 7Æ5 min respectively (data not
shown). After the required incubation time, 1 ml of the
bacterial suspension was removed and added to 9 ml of a
triclosan solution in sterile distilled water and the lethal
activity was determined as described above. For the test
carried out with bacteria in stationary phase, 1 ml of an
overnight culture was added to 9 ml of a triclosan solution
prepared in sterile distilled water.
The neutralizer used here was based on the British
Standard EN 1499 (CEN 1997b). The solution was made
fresh in distilled water and contained 30 g l)1 polysorbate 80
(Tween 80; Sigma), 3 g l)1 lecithin from eggs (Sigma),
1 g l)1 L-histidine (Sigma), 5 g l)1 sodium thiosulfate
(Fisher). Both the efficacy and the toxicity of the neutralizer
478 M . G . E S C A L A D A ET AL.

were investigated prior to testing with methods described by
Walsh et al. (1999) and Langsrud and Sundheim (1998)
respectively.
A control to determine the activity of the highest
concentration of DMSO used in these tests was also
included, although only 15 s and 20 min contact time were
investigated.
Statistical analysis
Parametric and nonparametric analysis of variance was
carried out using Minitab Release 13 software (Minitab
Inc., Philadelphia, PA, USA). Assumptions of normal
distribution, equal variances and independent and normally
distributed residuals were confirmed using the Minitab
software. If violated, the Kurskal–Wallis test was performed
at 95% confidence interval, otherwise analysis of variance
(ANOVA) was performed at the same confidence interval.
RESULTS
Bacteria–triclosan interactions at low
concentrations
The MIC values of triclosan against the different standard
strains tested in broth and in agar, respectively, were as
follows: E. coli, 0Æ1 and 0Æ09–0Æ1 mg l)1; Ent. hirae, 5Æ0–5Æ5
and 5Æ0 mg l)1; S. aureus, 0Æ01 and 0Æ1 mg l)1. These results
are consistent with MIC values from the manufacturer
(Anon 1998), which noticed some variability in MICs among
not only different strains of E. coli but also other organisms
such as S. aureus. It is also interesting to point out that for
S. aureus there was a 10-fold difference in MIC whether
they were determined in agar or in broth.
The results of the effects of subinhibitory concentrations
of triclosan on the growth of E. coli are presented in Fig. 1.
Similar observations were made with the other two strains
tested and therefore, respective graphs are not presented.
The effects of DMSO on the duration of lag phase, slope of
the exponential phase and final OD540 in stationary phase
were not significantly different (P > 0Æ05) from the values
obtained for the growth controls. The addition of triclosan
to the growth media increased the lag phase in a concen-
tration dependent manner (R2 values: 0Æ9205, 0Æ9303 and
0Æ9766 for E. coli, Ent. hirae and S. aureus, respectively; data
not shown). Subinhibitory concentrations of triclosan signi-
ficantly affected (P < 0Æ05) the slope of the exponential
phase and hence the growth rate, although this effect was
not concentration-dependent (Fig. 1). There was a signifi-
cant difference (P < 0Æ05) in result between the control and
all the concentrations tested, but not between concentrations
themselves (P > 0Æ05). The final OD540 readings of treated
cultures were not significantly different from untreated
cultures (controls;Fig. 1), apart from concentrations above
the MIC (i.e. >0Æ1 mg l)1 for E. coli; Fig. 1) for which, no
growth was recorded.
Bacteria–triclosan interaction at high
concentrations: lethal effects
The co-solvent (DMSO) at the concentrations used did not
have a significant effect (P > 0Æ05) on the viability of the

1·4

1·2

1
OD (540 nm)

0·8

0·6

0·4

0·2

0
0 200 400 600 800 1000 1200 1400 1600
–0·2
Time (min)

Fig. 1 Effect of a range of triclosan concentrations on the phases of population growth of Escherichia coli in tryptone soya broth at 37
C. Triclosan
concentrations: (d) 0Æ02 lg ml)1, (h) 0Æ04 lg ml)1, (n) 0Æ06 lg ml)1, (s) 0Æ08 lg ml)1, (·) 0Æ1 lg ml)1, (+) 0Æ5 lg ml)1, ( ) 1% v/v
dimethylsulfoxide and ( ) growth control. Error bars indicating SD of three independent experiments have been removed for clarity

ª 2005 The Society for Applied Microbiology, Letters in Applied Microbiology, 41, 476–481, doi:10.1111/j.1472-765X.2005.01790.x
 TRICLOSAN INTERACTIONS WITH BACTERIA 479

organisms (data not shown). The lethal effects of triclosan
against the different strains tested at different phases of
population growth are summarized in Tables 1–3. Signifi-
cant differences in results between the different phases of
population growth were observed and were concentration
and contact time dependent. Overall, triclosan at high
concentrations was bactericidal against all micro-organisms
regardless of their phase of population growth. However,
some differences in inactivation were observed at lower
concentrations, particularly, for population in the stationary
phase for which longer exposure were needed to achieve at
least a 5 log10 reduction in number. In addition, washed

Table 1 Activity of triclosan against Escherichia coli at different phases of population growth

Triclosan concentration (mg l)1)

35 40 45 50

Population phase CT LR CT LR CT LR CT LR

Lag phase 2 6Æ57 ± 2Æ33 2 5Æ18 ± 4Æ73 2 7Æ92 ± 0Æ02 2 7Æ92 ± 0Æ02
Log phase 2 6Æ45 ± 2Æ65 2 6Æ74 ± 2Æ19 2 6Æ64 ± 2Æ37 2 6Æ72 ± 2Æ23
Stationary phase 10 5Æ46 ± 2Æ32 5 5Æ27 ± 2Æ55 10 5Æ60 ± 2Æ22 5 6Æ71 ± 2Æ50
Washed cells 20 2Æ83 ± 0Æ56* 20 1Æ71 ± 0Æ60* 20 3Æ30 ± 0Æ84* 20 2Æ86 ± 0Æ85*

Data shown represent the mean of at least three independent experiments.

CT, time taken to obtain 5 log10 reduction in bacterial count; LR, respective log10 reduction ± SD.
*The bisphenol failed to produce >5 log10 reduction after the maximum contact time exposure tested, 20 min.

Table 2 Activity of triclosan against Enterococcus hirae at different phases of population growth

Triclosan concentration (mg l)1)

5 10 15 20

Population phase CT LR CT LR CT LR CT LR

Lag phase 20 )0Æ01 ± 0Æ18* 20 2Æ75 ± 0Æ93* 5 5Æ38 ± 2Æ39 0Æ25 4Æ47 ± 2Æ78
Log phase 20 0Æ26 ± 0Æ43* 10 5Æ08 ± 2Æ68 2 6Æ68 ± 1Æ92 0Æ25 6Æ25 ± 2Æ20
Stationary phase 20 0Æ39 ± 0Æ48* 15 5Æ10 ± 2Æ63 2 4Æ72 ± 2Æ65 2 7Æ07 ± 1Æ88
Washed cells 20 0Æ69 ± 2Æ02* 20 0Æ21 ± 0Æ16* 20 2Æ71 ± 0Æ00* 20 3Æ95 ± 0Æ00*

Data shown represent the mean of at least three independent experiments.

CT, time taken to obtain 5 log10 reduction in bacterial count; LR, respective log10 reduction ± SD.
*The bisphenol failed to produce >5 log10 reduction after the maximum contact time exposure tested, 20 min.
‘)’, an increase in cell number.

Table 3 Activity of triclosan against Staphylococcus aureus at different phases of population growth

Triclosan concentration (mg l)1)

20 25 30 35

Population phase CT LR CT LR CT LR CT LR

Lag phase 20 5Æ00 ± 0Æ00 2 5Æ18 ± 2Æ30 2 8Æ03 ± 0Æ09 0Æ25 4Æ95 ± 2Æ84
Log phase 20 4Æ73 ± 0Æ47* 2 6Æ80 ± 2Æ43 2 6Æ95 ± 2Æ02 2 8Æ11 ± 0Æ19
Stationary phase 20 3Æ93 ± 0Æ95* 15 5Æ10 ± 2Æ63 5 5Æ38 ± 2Æ39 2 6Æ58 ± 2Æ72
Washed cells 20 2Æ51 ± 2Æ57* 20 3Æ29 ± 2Æ97* 2 4Æ79 ± 2Æ27 10 5Æ89 ± 2Æ36

Data shown represent the mean of at least three independent experiments.

CT, time taken to obtain 5 log10 reduction in bacterial count; LR, respective log10 reduction ± SD.
*The bisphenol failed to produce >5 log10 reduction after the maximum contact time exposure tested, 20 min.
‘)’, an increase in cell number.

ª 2005 The Society for Applied Microbiology, Letters in Applied Microbiology, 41, 476–481, doi:10.1111/j.1472-765X.2005.01790.x
480 M . G . E S C A L A D A ET AL.
bacterial suspensions were less susceptible to the bactericidal
effect of triclosan regardless of the test bacteria
(Tables 1–3).
Triclosan was more effective (P < 0Æ05) against E. coli in
its lag and log phase, and less effective against washed cells.
The differences in E. coli inactivation depending upon its
phase of population growth was contact time dependent but
not concentration dependent (Table 1). Similar observations
were made with Ent. hirae, whereby bacterial inactivation at
different phases of population growth became significantly
different (P < 0Æ05) with increasing exposure time, but also
with increasing concentrations, i.e. with triclosan concentra-
tions of 10 mg l)1 or above (Table 2). The bactericidal
activity of triclosan at different phases of population growth
was less pronounced with S. aureus although it was
dependent upon exposure time (Table 3). As with the other
two bacteria, washed cells were less affected by the
bisphenol.
DISCUSSION
The triclosan inhibitory results presented here are consistent
with the literature. Although MIC testing is particularly
useful for estimating antibiotic activity, it is generally less
appropriate with biocidal agents (Russell 2003). However,
MIC values of triclosan have been widely used to indicate
microbial increase insusceptibility to the bisphenol, partic-
ularly when the effects of altered target sites or multidrug
efflux pumps were studied (Heath et al. 1998, 1999, 2001;
McMurry et al. 1998a,b, 1999; Levy et al. 1999).
Here, subinhibitory concentrations of the triclosan had a
dramatic concentration- dependent effect on the duration of
the lag phase for three bacterial genera tested. These low
concentrations also affected their kinetic of growth as
exemplified by a decrease of the slope of the exponential
phase. This is particularly remarkable as phenolics in
general have a high concentration exponent (g > 6)
(McDonnell and Russell 1999), which means that their
activity is rapidly lost upon dilution. The delay in the onset
of growth caused by exposure to triclosan can only be
explained by a strong interaction of the bisphenol with the
cell and cell target(s). It is conceivable that triclosan induces
a stress response followed by, or in addition to, the
expression of mechanisms that reduce the deleterious effect
of the biocide, for example, efflux pumps (McMurry et al.
1998b; Gilbert et al. 2002). At higher concentrations,
triclosan was both rapid-acting and active at all phases of
population growth, although some marked differences were
observed in its lethality. These results substantiated earlier
findings with S. aureus (Regos and Hitz 1974; Suller and
Russell 2000). The enoyl reductase has been claimed to be
the sole target of triclosan (Heath et al. 1998; McMurry
et al. 1998a; Larkin 1999; Levy et al. 1999). However,
ª 2005 The Society for Applied Microbiology, Letters in Applied Microbiology, 41, 476–481, doi:10.1111/j.1472-765X.2005.01790.x
recent findings suggested that the inhibition of the fatty acid
biosynthesis might be involved in the growth inhibitory
action, but other mechanisms are also involved in its lethal
effect (Gomez Escalada et al. 2005). Our results corroborate
these latest observations, whereby at low concentrations,
bacterial growth was severely affected, but at high concen-
tration (i.e. here the highest concentration tested) the
bactericidal effect was rapid, indicating a more damaging
effect such as membrane activity (Villalain et al. 2001).
Although triclosan lethality was rapid against metabolically
active bacteria (i.e. in lag and log-population phase) its
efficacy was slower against stationary phase population and
even less so against washed bacterial suspension. These
results indicate that (i) metabolically inactive bacteria
(washed suspension) are more resilient to the lethal effect
of the triclosan and (ii) bioavailability of the bisphenol is
important. In the first case, the inhibition of key metabolic
pathway and synthesis (Regos and Hitz 1974; McMurry
et al. 1998a) might be part of the lethal action mechanisms
of the bisphenol. As for the second remark, it is likely that
samples from bacterial population in stationary phase
contained bacterial debris and dead bacteria, which might
contribute to the ‘mopping up’ of triclosan from the
solution, hence, decreasing its bioavailability. Although this
hypothesis was not confirmed, bioavailability of triclosan is
important for its activity (Gomez Escalada et al. 2005).
In conclusion, the interaction of triclosan with the
bacterial cell is complex and multifactorial. While the
inhibition of the key metabolic pathway and of the synthesis
machinery is likely at very low concentrations, its lethal
activity results from a combination of the latter with other
mechanisms, such as membrane activity, which becomes
predominant with increasing concentrations.
ACKNOWLEDGEMENTS
The authors would like to thank Ciba Specialty Chemicals,
Grenzach-Wyhlen, Germany for funding this investigation,
including a research studentship to MGE.
REFERENCES
Anon. (1998) General Information on Chemical, Physical and Micro-
biological Properties. Grenzach-Wyhlen, Germany: Ciba Specialty
Chemicals. Irgasan DP300/Irgacare MP/Irgacide LP10.
Bhargava, H.N. and Leonard, P.A. (1996) Triclosan: applications and
safety. Am J Infect Control 24, 209–218.
Brady, L.M., Thomson, M., Palmer, M.A. and Harkness, J.L. (1990)
Successful control of endemic MRSA in a cardiothoracic surgical
unit. Med J Aus 152, 240–243.
CEN (Comité Européen de Normalisation, European Committee for
Standardization). (1997a). EN 1276 Chemical disinfectants and
antiseptics – Quantitative suspension test for the evaluation of

bactericidal activity of chemical disinfectants and antiseptics for use in
food, industrial, domestic and institutional areas – Test method and
requirements (phase 2, step 1). London: British Standard Institute.
CEN (Comité Européen de Normalisation, European Committee for
Standardization). (1997b). EN 1499 Chemical disinfectants and
antiseptics – Hygienic handwash – Test method and requirements (phase
2, step 2). London: British Standard Institute.
Gilbert, P., Allison, D.G. and McBain, A.J. (2002) Biofilms in vitro and
in vivo: do singular mechanisms imply cross-resistance? J Appl
Microbiol 92 (Suppl. 1), 98–110.
Gomez Escalada, M., Harwood, J.L., Maillard, J.-Y. and Ochs, D. (2005)
Triclosan inhibition of fatty acid synthesis and its effect on growth of
E. coli and Ps. aeruginosa. J Antimicrob Chemother 55, 879–882.
Heath, R.J., Yu, Y.-T., Shapiro, S., Olson, E. and Rock, C.O. (1998)
Broad spectrum antimicrobial biocides target the FabI component of
fatty acid biosynthesis. J Biol Chem 273, 30316–30320.
Heath, R., Rubin, J., Holland, D., Zhang, E.L., Snow, M.E. and Rock,
C.O. (1999) Mechanism of triclosan inhibition of bacterial fatty acid
biosynthesis. J Biol Chem 274, 11110–11114.
Heath, R.J., White, S.W. and Rock, C.O. (2001) Lipid biosynthesis as a
target for antibacterial agents. Prog Lipid Res 40, 467–497.
Jones, R.D., Jampani, H.B., Newman, J.L. et al. (2000) Triclosan: a
review of effectiveness and safety in health care settings. Am J Infect
Control 28, 184–196.
Langsrud, S. and Sundheim, G. (1998) Factors influencing a
suspension test method for antimicrobial activity of disinfectants.
J Appl Microbiol 85, 1006–1012.
Larkin, M. (1999) A close look at triclosan raises questions. Lancet 353,
1160.
Levy, C.W., Roujeinikova, A., Sedelnikova, S., Baker, P.J., Stuitje, R.,
Slabast, A.R., Rice, D.W. and Rafferty, J.B. (1999) Molecular basis
of triclosan activity. Nature 398, 384–385.
McDonnell, G. and Russell, A.D. (1999) Antiseptics and disinfectants:
activity, action and resistance. Clin Microbiol Rev 12, 147–179.
ª 2005 The Society for Applied Microbiology, Letters in Applied Microbiology, 41, 476–481, doi:10.1111/j.1472-765X.2005.01790.x
TRICLOSAN INTERACTIONS WITH BACTERIA 481
McMurry, L.M., Oethinger, M. and Levy, S.B. (1998a) Triclosan
targets lipid synthesis. Nature 394, 531–532.
McMurry, L.M., Oethinger, M. and Levy, S.B. (1998b) Overexpres-
sion of marA, soxS, or acrAB produces resistance to triclosan in
laboratory and clinical strains of Escherichia coli. FEMS Microbiol
Lett 166, 305–309.
McMurry, L.M., McDermot, P.F. and Levy, S.B. (1999) Genetic
evidence that InhA of Mycobacterium smegmatis is a target for
triclosan. Antimicrob Agents Chemother 43, 711–713.
Regos, J. and Hitz, H.R. (1974) Investigations on the mode of action of
triclosan, a broad spectrum antimicrobial agent. Zentralbl Bakteriol
Hyg I Abt Orig 226, 390–401.
Russell, A.D. (2003) Biocide usage and antibiotic resistance: the
relevance of laboratory findings to clinical and environmental
situations. Lancet Infect Dis 3, 794–803.
Russell, A.D. (2004) Whither triclosan? J Antimicrob Chemother 53,
693–695.
Suller, M.T.E. and Russell, A.D. (2000) Triclosan and antibiotic
resistance in Staphylococcus aureus. J Antimicrob Chemother 46, 11–
18.
Villalain, J., Mateo, C.R., Aranda, F.J., Shapiro, S. and Micol, V.
(2001) Membranotropic effects of the antibacterial agent triclosan.
Arch Biochem Biophys 390, 128–136.
Walsh, S.E., Maillard, J.-Y. and Russell, A.D. (1999) ortho-Phthalal-
dehyde: a possible alternative to glutaraldehyde for high-level
disinfection. J Appl Microbiol 86, 1039–1046.
Webster, J., Foagali, J.L. and Cartwright, D. (1994) Elimination of
methicillin-resistant Staphylococcus aureus from a neonatal intensive
care unit after hand washing with triclosan. J Paediatr Child Health
30, 59–64.
Zafar, A.B., Butler, R.C., Reese, D.J., Gaydos, L.A. and Mennonna,
P.A. (1995) Use of 0.3% triclosan (Bacti-Stat*) to eradicate an
outbreak of methicillin-resistant Staphylococcus aureus in a neonatal
nursery. Am J Infect Control 23, 200–208.