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Elite Research Journal of Biotechnology and Microbiology Vol. 2(1) pp. 1 - 3, August, 2014
Available online http://www.eliteresearchjournals.org/erjbm/index.htm
Copyright © 2014 Elite Research Journals
Full Length Research Paper
SENSITIVITY PROFILE OF BACTERIAL FLORA ISOLATED
FROM BATHROOM
1
2
Ajayi A *. and Ekozien M. I.
Department of Microbiology, University of Lagos, Akoka, Lagos, Nigeria
2
Department of Microbiology, Ambrose Alli University Ekpoma Nigeria
1
Accepted 12 July, 2014
Several bacterial pathogens were isolated from the floor, door-handle and wall of bathrooms in four
male and four female hostels. Seven isolated bacteria were areStaphylococcus aureus,
Streptococcus pneumoniae, Bacillus subtilis, Enterococcus faecalis, Klebsiella pneumoniae,
Escherichia coli and Proteus mirablis. Population density of bacteria was more on walls compared
to other points analyzed. Most of the isolates were resistant to an array of antibiotics used for
susceptibility test. Gram positive bacterial isolates obtained were more resistant toGentamycin,
Augumentin (Amoxicillin Clavulanate) and Septrin (Co-trimoxazole). However some of these isolates
were highly susceptible to Ciproxin, while most Gram positive bacterial isolates exhibited partial
susceptibility.
Keywords: Resistance, Susceptibility, Antibiotics, Zone of inhibition
INTRODUCTION
Sanitary conditions in public places have always been a
major problem, especially in bathrooms. Health
departments are continually checking the cleanliness
and safety of these bacteria breeding places to prevent
thespread of
sickness and disease (Chris et al.,
2002).Data on public washrooms contamination shows
how often and easily high-contact washroom surfaces
can be contaminated. Sherifa (2013) reported the high
presence of bacteria in public female restroom at Taif
Kingdom of Saudi Arabia (KSA). Although people may
claim their personal hygiene and clean toilet facilities, a
number of researches have shown different picture
(Gerba, 1995). Bathroom is any building or room is
made for people to have their bath, usually with soap
and water. Most bathrooms comprise of integrated toilet
facilities and sinks for other related washings. Droplets
from flushing contain pathogen including faecal matter
and aerosol, due to the mechanism of flushing these
aerosol and faecal matter splatter onto floor, partitions,
walls and air (Baker and Bloomfield, 2000). The public
bathroom is suspected to have the highest
concentration and variety of bacteria because of their
use by a huge number of people.
Antibiotic resistant microorganisms (ARM) are as much
prevalent today in long-term care facilities as are in
acute care facilities. These microorganisms include
methicillin resistant Staphylococcus aureus(MRSA),
aminoglycoside resistant Gram negative Bacilli such as
Pseudomonas aeruginosa and emerging vancomycin
resistant Enterococci (VRE).
Corresponding Author E-mail: [email protected]
The widespread use of antibiotics is strongly associated
with the development of antibiotic resistantbacteria
(Bennett and Brachman, 1992).This study was
conducted to determine the presence of pathogenic
bacteria that abound in bathroom environment and to
determine their resistance to common antibiotics.
MATERIALS AND METHODS
SAMPLE COLLECTION:
Samples were obtained from bathrooms of four male
and four female hostels in Ambrose Alli University
Ekpoma Nigeria. Sterile swab sticks were used to
obtain samples from three points each (door-handle,
wall and floor) of bathrooms. Samples were
immediately transported to the laboratory for analysis.
MICROBIOLOGICAL ANALYSIS:
Swab sticks were dipped into 10 ml sterile peptone
water. 1 ml of each stock was serially diluted into 9ml of
sterile normal saline, from which 0.1 ml aliquots of
3
5
7
9
dilution 10 , 10 , 10 10 was inoculated into agar plates
o
and spread to dryness then incubated for 24 h 37 C.
(Bello, 2002)
ISOLATION AND IDENTIFICATION OF ISOLATES:
Samples were cultured on blood agar, nutrient agar,
and MacConkey agar plates (Oxoid, Basingstok,
England). Bacteria isolates obtained were subjected to
biochemical test for identification of pathogenic
bacteria.
Ajayi and Ekozien 2
ANTIBIOTIC SUSCEPTIBILITY TEST:
RESULTS
A light suspension (0.5 McFarland standard) of the test
isolates were poured onto the surface of freshly
prepared agar plates and the excess was gently
drained off. The agar surface was allowed to air dry,
and then the antibiotic discs were placed on the agar
surface using sterile forceps. Plates were overturned
0
and incubated at 35 C for 24 h. Zone of inhibition
measuring up to ≥ 20 mm indicated susceptibility
(Bello, 2002).
Enumeration of microbial populations:
Total bacteria counts revealed that population density of
bacteria varied from different sampling points (doorhandle, wall and floor). Bacteria population density on
5
11
walls ranged from 6.3×10 cfu/ml to 2.35×10 cfu/ml
9
compared to a maximum of 2.13×10 cfu/ml on floor of
8
female hosteland 1.12×10 cfu/ml on door handle of
male hostel.
Table 1: Average Population Densities of Bacteria Isolates on Sampling Points
Sampling Points
Floor
Door-handle
Wall
Bacteria population density (cfu/ml)
Male Hostel
Female Hostel
8
9
2.32×10
2.13×10
8
6
1.12×10
2.63×10
10
11
6.3×10
2.35×10
prevalent(43%) followed by Enterococcus faecalis(21%)
as shown in Table 2. Klebsiella pneumoniaeand
Bacillus subtilis showed the lowest percentage (1%) of
occurrence.
Characterization and identification of bacteria isolates:
The isolates were identified using colonial morphology,
and biochemical tests. Results showed isolates as
Staphylococcus aureus, Streptococcus pneumoniae,
Bacillus subtilis, Enterococcus faecalis, Klebsiella
pneumoniae, Escherichia coli and Proteus mirablis.
Staphylococcus aureuswas found to be the most
Table 2: Frequency of occurrence of bacteria isolates
Bacteria isolates
Staphylococcus aureus
Escherichia coli
Proteus mirablis
Enterococcus faecalis
Bacillus substilis
Klebsiella pneumonia
Streptococcus pneumonia
Frequency of occurrence/%
53
43%
41
12%
37
18%
43
21%
16
1%
16
1%
19
4%
Streptomycin with the exception of Bacillus subtilis
which was partially sensitive to Streptomycin as
indicated in Table 4. Streptococcus pneumoniae was
resistant to Ciprofloxacin and Enterococcus faecalis
was partially susceptible to Ciprofloxacin, while Bacillus
subtilisand Staphylococcus aureuswere susceptible to
Ciprofloxacin.
Antibiotic susceptibility test revealed that Gram negative
bacteria isolates were resistant to Zinnat (Cefuroxime
axetil) and Augumentin (Amoxicillin Clavulanate).
Though they were also resistant to Nalidixic acid and
Ampicillin Escherichia coli was partially susceptible to
both antibiotics Table 3. All Gram positive bacterial
isolates were resistant to Augumentin (Amoxicillin
Clavulanate), Cloxacillin, Septrin (Co-trimoxazole),
Ampicillin, Gentamycin and Erythromycin. All Gram
positive bacterial isolates were not susceptible to
Table 3: Susceptibility ofGram negative bacterial
isolates to antibiotics (zone of inhibition in mm)
Bacteria
Proteus mirablis
OFX
23
CXM
0
CPX
24
AU
0
CN
21
S
16
COL
21
NA
0
SXT
21
PN
0
Klebsiella pneumonia
24
2
24
1
14
12
15
5
22
2
Escherichia coli
21
0
26
2
15
0
1
16
2
14
≥ 20 = Susceptible,
≤ 11 = Resistant
3 Elite Res J. Bio M.
Table 4: Susceptibility ofGram Positive bacterial isolates to antibiotics (zone of inhibition in mm)
Bacteria
Staphylococcus aureus
Bacillus substilis
Enterococcus faecalis
Streptococcus pneumoniae
AU
0
0
0
0
CXL
0
0
0
0
SXT
0
0
0
0
CRO
3
21
0
0
OFX – Ofloxacin
CPX – Ciprofloxacin
CN – Gentamycin
COL- Colimycin
SXT – Septrin (Co-trimoxazole)
E – Erythromycin
CRO – Ceftriaxone
The zone of inhibition ranged from 1mm to 26 mm for
Gram negative bacteria and 1 mm to 22 mm for Gram
positive bacteria isolates.
DISCUSSION
Except for an operating room or other sterile
environments, bacteria can be found almost
everywhere, which is because they are very resilient.
Public bathrooms are the perfect place for bacteria to
strive because these facilities are designed to dispose
of large volume of human waste, which consist of 30%
bacteria (Cassidy, 1987). This study corroborates this
fact as high population densities of bacteria were
recorded from the different places of bathrooms
sampled. Chris et al., (2002) also reported the presence
of bacteria isolates such as Staphylococcus aureus,
Escherichia coli and Salmonellaspp. from the
bathrooms of students at the university of Miami USA.
Also Opere et al., (2013) reported the isolation of
Bacillus spp., Staphylococcus aureus, Staphylococcus
epidermidis, Micrococcus spp., Pseudomonas spp.,
Enterococcus faecalis, Salmonella typhi and Shigella
dysenteriae with varying resistance to a range of
antibiotics from public toilets.
The presence of organisms such as Staphylococcus
aureus, Klebsiella pneumoinae, Escherichia coli,
Streptococcus pneumonia and Salmonellaspp. have
been reported to have cause diseases which is a
source of concern. Though according to Bennett and
Brachmen (1992) the role of the environment in the
transmission of antibiotic resistant microorganism(ARM)
has not been well established.Results of this study
revealed that antibiotic resistant microorganism
contaminates environmental surfaces such as
bathrooms. Most isolates obtained in this study were
resistant to most commonly used antibiotics such as
Septrin, Agumentin, Cloxacillin, Gentamycin and
Ampicillin.This concurs with the findings of Adewoyin et
al., (2013) and this may be attributed to misuse of
antibiotics by students.
PN
0
0
1
0
S
5
16
2
0
CN
3
4
2
0
CTX
0
20
0
0
E
0
0
0
0
CPX
19
20
16
0
CXM – Zinnat (Cefuroxime axetil)
AU – Augumentin
S- Streptomycin
NA – Nalidixilic acid
PN – Ampicillin
CTX – Cefotaxime
CXL – Cloxacillin
CONCLUSION
The presence of antibiotic resistant bacteria (ARB) in
the bathrooms of student is a matter of concern, since
they generally pose health risk to individuals. This
implies that transmission of ARB through bathrooms is
possible; therefore proper sanitary practice should be
adopted by all where necessary to prevent the spread
of pathogenic bacteria.
REFERENCES
Ajibade, V. A., Aboloma, R . I and Oyebode, J. A. (2010).
Survival and Antimicrobial Resistance of Salmonella
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Adewoyin A. G., Majolagbe, O. N., Ogunmoyewa, T. and
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Microbial Isolates of Toilet-Bowl of some Students
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nd
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rd
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Cassidy, A. (1987). Bathroom infection. Journal of Hospital
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Chris, J., James, D., Paul, G., and Michelle, C. (2002). The
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Gerba, C. (1995). Microorganisms in Public Washrooms.
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Opere B. O, Ojo J. O, Omonighehin E. and Bamidele M.
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