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Antibacterial activity of soaps against daily encountered bacteria

Article  in  AFRICAN JOURNAL OF BIOTECHNOLOGY · April 2009

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African Journal of Biotechnology Vol. 8 (8), pp. 1431-1436, 20 April, 2009
Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 © 2009 Academic Journals

Full Length Research Paper

Antibacterial activity of soaps against daily

encountered bacteria
Saba Riaz*, Adeel Ahmad and Shahida Hasnain
Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore 54590, Pakistan.
Accepted 6 February, 2009

This study aims to check the antibacterial activity of various branded soaps against bacteria that are
normally present in the environment. The proposed study includes selection of most common bacterial
strains from the environment. Identification of bacterial strains was done by standard microbiological
techniques, which include gram staining, biochemical testing and advanced identification by analytical
profile index. Determination of minimal inhibitory concentration and minimum bactericidal activity of
strains was performed by tube and microtitration method. Antibacterial soaps showed better MIC in
comparison with beauty soaps. The most resistant bacterium to all the soaps is Klebsiella pneumoniae
and Pseudomonas aeruginosa. It is obvious that antibacterial soaps have the antibacterial agents that
can either kill or inhibit the bacterial cells. It might be possible that some bacterial strains become
resistant which leads to their survival even at high concentrations of soaps.

Key word: Soaps, antibacterial activity, daily bacteria.

INTRODUCTION

Soaps play an important role in removing and killing tion has been well recognized (HWG, 1999). The
bacteria. Although fats and oils are general ingredient of importance of hand hygiene is also there for food
soaps but some detergents are added to enhance the handlers. Food handler includes those who deals with
antibacterial activities of soaps (Friedman and Wolf, delivers and serve food (Horton and Parker, 2002). Dr
1996). According to Osbore and Grobe antibacterial Elaine Larson published an extensive review and
soaps can remove 65 to 85% bacteria from human skin concluded that hand hygiene and reduced tran-smission
(Osborne and Grube, 1982). Bacteria are very diverse of infections is a convincing fact (Larson et al., 1986). To
and present every where such as in soil, water, sewage, investigate the antibacterial efficiency of different brands
standing water and even in human body. Bacteria’s that of soaps, we isolated bacteria from different environ-
attacks on human body is of great importance with ments and human skin. Identification of bacteria was
reference to health (Johnson et al., 2002). Transient done by biochemical tests (Cheesbrough, 2001) and by
bacteria are deposited on the skin surface from environ- using analytical profile index. Minimum inhibitory concen-
mental sources and causes skin infections. Examples of tration (MIC) and minimum bactericidal concentration
such bacteria are Pseudomonas aeruginosa (Fluit et al., (MBC) against these bacteria were determined. Identifi-
2001) and Staphylococcus aureus (Higaki et al., 2000). cation of bacterial species that are most resistant to the
The importance of hand washing is more crucial when it antibacterial soaps of daily use was made. The present
is associated to health care workers because of possible studies were aimed to determine the bactericidal
cross contamination of bacteria that may be pathogenic activity/efficacy of both the antibacterial as well as beauty
or opportunistic (Richards et al., 1995). Studies have soaps and to determine, whether the soaps only removes
shown that soaps containing antimicrobial active ingre- the bacteria from skin or it also kills the bacteria.
dients remove more bacteria as compared to plain soap
(Lucet et al., 2002). Handhygiene and prevention of infec-
MATERIALS AND METHODS

Soaps used

*Corresponding author. E-mail: [email protected]. Safeguard soap, Lifebuoy Red soap, Lifebuoy White soap, Dettol
1432 Afr. J. Biotechnol.

Table 1. Sampling sites of Bacteria.

S/N Bacteria Collection area

1 Staphylococcus aureus Soil
2 Pseudomonas aeroginosa Waste Water
3 Escherichia coli Sewage Water
4 Pseudomonas Waste Water
5 Klebsiella pneumoniae Polluted Soil
6 Klebsiella pneumoniae Polluted Soil
7 Enterobacter spp. Sewage Water
8 Pseudomonas Waste Water
9 Bacillus subtilis Under Nails
10 Staphylococcus epidermidis Skin

bar soap, Dettol Liquid hand wash (antibacterial soaps), Lux soap, Safeguard soap MBC is 250 mg/ml and its MIC was
Palmolive soap, and Capri soap (Beauty soaps). Safeguard is an antibacterial soap that has bactericidal
observed at 125 mg/ml for Staphylococcus aureus.
Isolation of bacteria Safeguard is an antibacterial soap that has bacteric
idalagents in it. For Pseudomonas aeroginosa (1) its
Different bacterial strains were isolated (Table 1). All samples were MBC was at 500 mg/ml and MIC was at 250 mg/ml. If it is
properly diluted and spread on the nutrient agar. The pH was compared with the S. aureus it clearly showed that it was
adjusted to 7.0, incubated for 24 h at 37°C. Then inoculation was killed at high concentration of soaps. The MBC
made on nutrient agar plates and incubated at 37°C for 24 h. The
most abundant strain of the samples was selected, gram stained,
ofsafeguard soap against strain of P. aeroginosa strain 2
and then purified on new plate of nutrient agar. was observed at 250 mg/ml and its MIC was observed at
25 mg/ml. For E. coli MBC was 125 mg/ml and its MIC
was observed at 62.5 mg/ml. Safeguard soap was used
Identification of isolated bacteria against Klebsiella pneumoniae its MBC at 500 mg/ml and
Identification of bacteria was done by using different biochemical
its MIC was observed at 250 mg/ml. The MBC of
tests. These tests were based on the gram stain reaction of safeguard soap against this strain of Pseudomonas
bacterial strains. Tests includes, oxidase test, catalase test, wasobserved at 250 mg/ml and its MIC was observed at
urease test, motility test, acid production from glucose, mannitol, 125mg/ml (Figure 3). If it is compared with the first strain
sucrose, lactose, maltose, coagulase test, Dnase test, indole test, of P. aeroginosa two prominent differences were
eosine methylene blue test, triple sugar iron reactions,
observed which showed that strain was more sensitive
methyl red test, voges proskauer test, and nitrate reduction test
following chesseborugh (Cheesbrough, 2001). For confirmation of than the first one. Dettol soap showed MBC at 250 mg/ml
gram negative bacteria, analytical profile index (biomereux) was and MIC was observed at 125 mg/ml against S. aureus.
performed according to manual instructions (Table 2). These values were compared to the values obtained from
safeguard soap. These were almost equal to the values
Strain maintenance
of safeguard. It might be estimated that the antibacterial
activity of safeguard and dettol soap were almost the
All strains were grown on nutrient agar plates at 37°C for 48 h. same against this organism. Lifebuoy red (antibacterial):
Strains were stored at -70°C in 50% sterile glycerol and TSB (Aulet, This soap showed its MBC at 250 mg/ml and MIC was
2001). Minimum inhibitory and bactericidal concentrations of observed at 125 mg/ml against S. aureus. The
different soaps To determine minimum inhibitory concentrations of comparison of safeguard, lifebuoy and dettol soaps
different soaps, two different methods were used. One was tube
method (Cappuccino and Sherman, 1992) and other was micro
revealed the equivalence of MBC and MIC values. It was
titration plate method (Johnson et al., 2002). Bactericidal also estimated that the organism might be sensitive to the
concentrations of soaps were determined following Cappuccino and antibacterial-soaps.-Lifebuoy-(Red)-is-also--an tibacterial
Sherman (1992). soap, it showed its MBC against is Pseudomonas spp. At
350 mg/ml and MIC was seen at 175 mg/ml which much
lower than safeguard and dettol soap. The antibacterial
RESULTS
agents used in this soap showed more antibacterial
The present study suggested that the choice of soap activity in comparison with the above mentioned soaps.
should be that which does not affect the facial tissues as This soap showed its MBC at 250 mg/ml and MIC at 150
well as effective against disease causing bacteria in a mg/ml or close to it. For Escherichia coli and Klebsiella,
small amount. For the determination of MBC and MIC, this soap showed its MBC at 500 mg/ml and MIC was
soaps of daily use were employed (Figures 1 and 2). seen at 250 mg/ml which is very high concentration of the
 Riaz et al. 1433

0.45

0.4

0.35 Safeguard
Optical Density
Lifebuoy(Red)
0.3
Dettol
0.25 Lux
0.2 Palmolive
Dettol(Liquid)
0.15
Lifebuoy(White)
0.1 Capri

0.05

0
0 100 200 300 400 500 600 700 800
Concentration mg/ml

Figure 1. Antibacterial activity of different soaps against Pseudomonas aeroginosa.

0.5
0.45
0.4 Safeguard
0.35 Lifebuoy(Red)
Optical Density

0.3 Dettol
Lux
0.25
Palmolive
0.2 Dettol(Liquid)
0.15 Lifebuoy(White)

0.1 Capri

0.05
0
0 100 200 300 400 500 600 700 800
Concentration mg/ml

Figure 2. Antibacterial activity of different soaps against Klebsiella pneumoniae.

800
700
600
MBC mg/ml

500
400
300 MBC mg/ml
200
100
0
sa

is
e
s

37

pp
lla iae s

s
eu

id
ia

id ilis
9

on na

oc illu ona
no

pn EB
B

m
on

rs
as aur

ep bt
E

o
gi

er
m
m

om
te

s su
ro

o li

eu
us

ac
ae

.c

ud

cu s
om cc

ob
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E

se
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er
s

oc c
u
P

yl Ba
on

ne

nt
se ylo

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E
bs
ph

K
ud

le
ta

ph
S

ta
P

Bacterial Species

Figure 3. Safeguard soap activity against different bacteria species.

1434 Afr. J. Biotechnol.

800
700
600
MBC mg/ml 500
400 MBC mg/ml
300
200
100
0 Staphylococcus

Staphylococcus
E.coli EB9

K.pneumoniaeEB37

pneumoniae
Pseudomonas

Pseudomonas

Pseudomonas
Enterobacter spp

Bacillus subtilis
Klebsiella
aeroginosa

epidermidis
aureus

Bacterial Species

Figure 4. Lifebuoy (White) soap activity against different bacteria species.

Table 2. Characteristics of the bacterial strains.

Bacteria
Test 1 2 3 4 5 6 7 8 9 10
Oxidase NA +ve -ve +ve -ve -ve -ve +ve +ve -ve
Catalase +ve +ve +ve +ve +ve +ve +ve +ve +ve +ve
Motility NA +ve +ve +ve -ve -ve +ve +ve +ve -ve
Lactose NA +ve +ve +ve +ve +ve +ve +ve +ve +ve
EMB NA NA +ve NA -ve -ve -ve NA NA NA
Indole NA -ve +ve -ve -ve -ve -ve -ve NA NA
Citrate NA +ve -ve +ve +ve +ve +ve +ve +ve NA
VP NA -ve -ve -ve +ve +ve +ve -ve NA NA
MR NA +ve +ve +ve -ve -ve -ve +ve NA NA
TSI NA R/R/-/- Y/Y/+/- R/R/-/- Y/Y/+/- Y/Y/+/- Y/Y/+/- R/R/-/- NA NA
Urease NA -ve -ve -ve +ve +ve -ve -ve NA +ve
Mannitol +ve +ve NA +ve NA NA NA +ve NA -ve
Maltose NA -ve -ve -ve -ve -ve -ve -ve -ve +ve
Pigment Golden Green -ve Blue -ve -ve -ve Blue -ve -ve
Coagulase +ve NA NA NA NA NA NA NA NA -ve
DNase +ve NA NA NA NA NA NA NA NA NA
Sucrose +ve -ve -ve -ve +ve +ve -ve -ve +ve +ve
Genera S.aureu P. aerog E. coli Ps spp K. neum K.pneu Enter p. P aerog B. ubtilis S. epid

soap. These bacterial spp. showed resistance to the soap bacterium but concentration was high as compared to the
at a very low concentration and were killed at very high antibacterial soap. The MBC was observed at 500 mg/ml
concentration. Staphylococcus epidermidis was killed at and MIC of the soap was 250 mg/ml. This revealed that
175 mg/ml that is very low concentration of soap and it Lux soap also showed antibacterial activity but not as
showed its MIC at 87.5 mg/ml, the antibacterial agent much, than the other specific antibacterial soaps. It might
proved to be efficient against this bacterium but this soap also be possible that S. aureus was sensitive to the Lux
also killed other bacteria like Enterobacter, B. subtilis at soap. This soap showed its MBC at 700 mg/ml and it also
350 mg/ml and there was no apparent growth observed showed its MIC at 350 mg/ml against Bacillus subtilis.
on the nutrient agar plate (Figure 4). Lux is a beauty soap This soap showed its MBC at 250 mg/ml and MIC at 125
and was used against S. aureus to check its antibacterial mg/ml against Pseudomonas aeroginosa 1 that was
activity. Lux contains Aloe vera which might have almost equal to the Lifebuoy red, it might be possible that
antibacterial activity. At 500 mg/ml, the Lux killed the some natural ingredients such as extract of A. vera show-
 Riaz et al. 1435

ed the antibacterial activity. The observation of Lux that plain soaps also possessed antibacterial activity
beauty soap revealed that these might posses’ germicidal although lesser than that of antibacterial soaps. Garner
activity. Palmolive beauty soap: It is beauty soap and was and Favero (1985) studied the hand washing with plain
used against S. aureus. Although this was not an soaps removes millions of microorganisms (Garner and
antibacterial soap but it showed it’s MBC at 500 mg/ml Favero, 1985). Most of the research has been focused on
and its MIC was observed at 250 mg/ml. This showed hand washing and hand disinfectants for personnel in
Palmolive soap can kill bacteria. At 500 mg/ml the health care settings where patients are immune
organism did not showed growth on the surface of the compromised and are at high risk. Bannan and Judge
nutrient agar medium. This soap showed its MBC at 700 (1965) indicated that hand washing with bar soap
mg/ml and MIC was observed at 350 mg/ml against reduced bacterial population (Bannan and Judge, 1965).
Bacillus subtilis this is equal or almost equal to the Lux Tierno (1999) response to the Association for Profess-
beauty soap but it is very high in comparison to the ionals in Infection Control and Epidemiology (APIC)
Lifebuoy both red and white this might be possible that emphasized the use of antimicrobial household products
due to lack of specific antibacterial agent it did not show (Tierno, 1999).
its MBC and MIC at low concentration. Lifebuoy white: It It was seen clearly that Gram positive bacteria were
also showed the MBC at 350 mg/ml and MIC at killed at low concentration of soaps than Gram negative
175mg/ml against B. subtilis. These values were equal or bacteria. The best of all the soaps used is lifebuoy white
almost equal to the Lifebuoy red soap and lower than the (antibacterial) because the calculation of the efficiency of
Dettol and Safeguard soap at 350 mg/ml there was all the soaps revealed that this soap is more efficient than
complete absence of bacterial growth on the agar plates the others used. The most resistant bacterium of all the
and at 175 mg/ml there was no growth. After 24 h of soaps is K. pneumoniae following P. aeruginosa. It is
incubation, few colonies were observed. The MBC of proved experimentally that antibacterial soaps kill the
soap against E. coli was observed at 125 mg/ml and MIC bacteria at a specific concentration; they also have bac-
was 62.5 mg/ml that is very low concentration of soap. teristatic activity and can inhibit the growth of bacteria.
The E. coli showed sensitivity for this antibacterial soap Beauty soaps contain some natural and plant extracted
as it was killed at very low concentration. The ingredients in their composition which have the ability to
antibacterial agent of Lifebuoy white soap might be inhibit or kill the bacteria so they also gave some
efficient in killing the cells. For Pseudomonas and K. bactericidal activity. Micro-titration plate method is
pneumoniae, the soap showed its MBC at 250 mg/ml and efficient than tube method and easier to perform. This
MIC was observed at 125 mg/ml. As this soap showed study suggests that selection of soaps should depend on
the MBC at 250 mg/ml and at this concentration no to the working environment. The soap should have good
growth of the bacteria was observed. So, the soap is ingredients which have the ability to kill bacteria but not to
efficient in killing the bacterium at this concentration. For damage body tissues. Health care workers should use
K. pneumoniae, Enterobacter spp. and B. subtilis, the soaps according to criteria of Health and Hygiene. In this
MBC was seen at 350 mg/ml and MIC was observed at way many immuno-compromised or low immunity
175 mg/ml. Lifebuoy showed its MBC for S. epidermidis patients can be protected from transfer of pathogenic or
at 700 mg/ml and MIC was seen at 350 mg/ml. The opportunistic pathogens. This area of research requires
organism was isolated from human skin and found Gram attention of scientists and people from soap industry,
positive but it was killed at 750 mg/ml concentration of because quality of soaps is very important as they are the
soap that was very high. need of every home.

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