San Pedro College

12 C. Guzman Street, Davao City,

Philippines

Bacterial Contamination of Disinfected and Non-disinfected Writing Pens

Used by Third Year Bachelor in Medical Laboratory Science Students

Curaza, et al.
Introduction
All students are potential carriers of potentially pathogenic or non-
pathogenic bacteria
Based on the WHO guidelines, good hand hygiene can lower the risk
of hand transmission of microorganisms
Detecting possible vectors of pathologic microorganisms is
another important step in blocking the transmission or
eradicating these pathogens.
The cleaning agents mostly used for removal of microorganisms
from the devices are alcohol swabs, but sterile water swabs are
also effective. However, manufacturers mainly recommend
(74%) cleaning the devices with water and detergent.
Recommendations to reduce contamination risks include staff
education, strict hand hygiene measures, and guidelines on
device cleaning.
PREVEN TION IS BETTER THAN CURE.
Problem
Most equipment harbor potential pathogens.
Students do not wash their hands after touching
contaminated objects thus transmitting potentially
pathogenic bacteria to other objects.
Potentially pathogenic bacteria could contaminate
writing pens posing a harmful threat to the students.
While other bacteria are mostly the normal flora of
the human body, it is still possible to harbor
pathogenic bacteria from writing pens.
Previous studies show that the most common bacteria
found on writing pens is the Micrococcus spp., which
is a skin commensal. Other common bacteria found
are Staphylococcus spp. and Enterococcus spp.
Hypothesis
Ho
There is no significant difference in the
bacterial growth of disinfected and non-
disinfected writing pens of third year BMLS
students.

Ha
There is a significant difference in the
bacterial growth of disinfected and non-
disinfected writing pens of third year BMLS
students.
Significance of the study
To identify and study the types of bacteria present in
the writing pens of the Bachelor of Medical Laboratory
Science students of San Pedro College.
To determine the frequency of contamination of
bacteria that contaminates the Disinfected and non-
disinfected writing pens of BMLS students of San
Pedro College.
To study and determine the other factors that affects
the transmission of bacteria.
To formulate and gather effective use of 70% Isopropyl
alcohol for the prevention of further transmission of
possible pathogenic bacteria that may cause disease
among BMLS students of San Pedro College.
Methods
SAMPLE SELECTION
Third year Bachelor in Medical Laboratory
Science students of San Pedro College
belonging to one section.

BACTERIAL SWAB
-sterile saline swab wiped on the surface of
the pens
-swab is performed on the pens used after
one laboratory meeting
Methods

NUTRIENT AGAR
Composed of: 0.5% peptone, 0.3% beef extract/
yeast extract, 1.5% agar, 0.5% NaCl, distilled
water
-general purpose, nutrient medium
-used for cultivation of microbes supporting
growth of a wide range of non-fastidious
organisms
-can grow a variety of types of fungi and
bacteria
Methods
GRAM STAINING
Gram staining separates bacteria into two
groups and uses the cell wall composition as
the basis of differentiation.
-Gram positive bacteria: stains purple
-Gram negative bacteria: stains red/ pink
Steps involved: Fixation, application of primary
stain (Crystal violet), application of mordant
(iodine), decolorization with the use of acetone
or ethanol, application of counterstain (safranin)
Methods
GRAM POSITIVE BACTERIA
Methods
GRAM NEGATIVE BACTERIA
Methods

OTHER TESTS:
o Catalase
o Coagulase
o Optochin disc sensitivity test ( Taxo-P):
differentiates Strep. Pneumoniae from alpha-
hemolytic streptococci.
Methods
OTHER TESTS

o Bacitracin disc: sensitivity test (Taxo A)

o Bile-esculin hydrolysis test (BEA): for Group D
Strep.
o 6.5 % NaCl (salt tolerance test) for group D
Streptococci (enterococcus)
Methods
OTHER TESTS

o CAMP ( Christie, Atkins, Munch, Peterson): for

group B strep (Streptococcus agalactiae)
o DNAse test: confirmatory test for identification
of staph aureus
o Germ tube
Methods
OTHER TESTS

o Biochemical tests
Methods

70% Isopropyl Alcohol

-one of the most commonly used antiseptics
by students for it is affordable and are very
available in supermarkets and other stores
-The effectivity of 70% Isopropyl Alcohol is
against the bacteria found on the pens will be
tested
Conceptual Framework

Positive Control:
Pens disinfected with 70% isopropyl alcohol.

Negative Control:
Pens that are not disinfected with 70% isopropyl alcohol

Independent Variable: 70% Isopropyl Alcohol

Dependent variable:
Bacterial growth on nutrient agar
RRL
A prospective study was done to assess the potential of writings pens as a fomite for hospital-acquired
pathogens. There was an incorporated intervention group that have pens that are wiped with alcohol-
based sanitizing agent between patient visits and organism load and bacteriological profile was
compared with the non-intervention group. Results show a reduced number of pens that showed visible
growth on culture and reduced Gram-positive cocci, bothStaphylococcusandEnterococcusspp on pens
wiped with alcohol-based sanitizing agent. The finding is essential for indicating that the risk of
transmission of fomites may be reduced when there is an application of alcohol based sanitizing agent
in wiping pens between patient encounter. (Halton et al. 2011)

A similar study was conducted in order to evaluate the role of pens in relation to transmission of
bacteria used by nursing students working in clinics. Results show that all the samples consisted of pens
and bacterial contamination was observed with a percentage of 12.5. The microorganisms identified
were: one sample (2.5%) wasStaphylococcus hominis, two samples (5%) wereStaphylococcus
epidermidis, one sample(2.5%) wasStaphylococcus haemolyticus, and one sample (2.5%)
wasStaphylococcus warneri. The study findings emphasizes the risks of nosocomial infections,
especially in pediatric units, if not enough attention is paid to the cleanliness of the pens used in the
clinics. (Cinar et al. 2014)

In 2009, a study was conducted to evaluate the role of pens as fomites used by respiratory therapists in
an intensive care unit.20 were obtained from respiratory therapists. Samples were cultured,
enumerated and identified. Result show that 17 out of 20 was positive the presence of bacteria, found
to have coagulase-negative staphylococci. The mean +/- SD number of colony-forming units was 126
+/- 277 with a range of 0-1,250. On 4 pens Micrococcus species were also found. Based on findings,
protocols used to reduce the transmission of pathogens may need to be extended to pens. (Wolfe et al.
2009)
RRL
70% isopropyl alcohol kills microorganism by protein denaturation or
dissolving the cell membrane of the cell wall. Bacteria with thin layer of
peptidoglycan can be easily destroyed by alcohol. Water acts plays a role in
denaturing the proteins of the cell membrane. It aids in the contact time
between the alcohol and microorganism. 70% alcohol gives a longer contact
time because the alcohol evaporation time is longer. (Choudhary, 2008)
The antimicrobial action of alcohol is denaturation of proteins. An absolute
ethyl alcohol, a dehydrating agent, is less bactericidal than alcohol mixed
with water because proteins are denatured more quickly in the presence of
water. (CDC)
Addition of water to alcohol breaks the hydrogen bonds between the alcohol
molecules and make molecules active. There is a critical concentration of
water molecules which makes a maximum free alcohol molecules. 30%
water concentration is critical concentration for isopropyl alcohol therefore,
70% isopropyl alcohol is used as disinfectant (Saxena, 2008)
Staphylococcus aureuskilled in less than 10 seconds with a 50% Isopropyl Alcohol
solution and was not killed in 2 hours with a 90% solution: emphasizing the importance
of water in killing microorganisms (Tainter et al.1944)