GUIDELINES OF DISSERTION

FOR B.Sc (MLT) DEGREE

DETECTION OF SURFACE CONTAMINATION ON COMPUTER

KEYBOARD AND MICE FROM DIFFERENT LOCATIONS OF

MOTHER HOSPITAL CAMPUS

DENNA FRANCIS

MOTHER COLLEGE OF PARAMEDICAL SCIENCES.THRISSUR

PROJECT SUBMITTED IN PARTIAL FULFILMENT OF

THE REQUIREMENTS FOR THE DEGREE OF BACHELOR

OF SCIENCE IN MEDICAL LABORATORY TECHNOLOGY

KERALA UNIVERSITY OF HEALTH SCIENCES

2015 – 2016
DETECTION OF SURFACE CONTAMINATION ON COMPUTER

KEYBOARD AND MICE FROM DIFFERENT LOCATION

OF MOTHER HOSPITAL CAMPUS

by

DENNA FRANCIS

Project Submitted to the

KERALA UNIVERSITY OF HEALTH

AND ALLIED SCIENCES THRISSUR

In partial fulfillment

Of the requirements for the degree of

B.Sc MLT

Under the guidance of

Mrs. SAJITHA S. NAIR.
Asst.Professor in Microbiology, Mother college of paramedical sciences

THRISSUR
 DECLARATION BY THE CANDIDATE

I DENNA FRANCIS hereby declare that thesis entitled –“DETECTION OF

SURFACE CONTAMINATION ON COMPUTER KEYBOARD AND MICE FROM

DIFFERENT LOCATION OF MOTHER HOSPITAL CAMPUS” is a bonafide and

genuine research work carried out by me under the guidance of Mrs.Sajitha S. Nair

Asst. professor in Microbiology, Mother college of paramedical science.

Signature of the candidate:

Place:
Date :
 CERTIFFICATE BY THE GUIDE

This is to certify that the dissertation entitled - DETECTION OF SURFACE

CONTAMINATION ON COMPUTER KEYBOARD AND MICE FROM DIFFERENT

LOCATION OF MOTHER HOSPITAL CAMPUS - is a bonafide research work done by

DENNA FRANCIS in partial fulfillment of the requirement for the degree of BSC MLT.

Signature of the Guide:

Name and Designation:

Place:

Date:
 ENDORSEMENT BY THE HOD

This is to certify that the project entitled – DETECTION OF SURFACE

CONTAMINATION ON COMPUTER KEYBOARD AND MICE FROM

DIFFFERENT LOCATION OF MOTHER HOSPITAL CAMPUS

is a bonafide research work done by DENNA FRANCIS partial fulfillment of the

requirement for the degree of BSC MLT.

Seal and Signature of HOD Seal and Signature of PRINCIPAL

Name: Name:

Place: Place:

Date Date:
 DECLARATION

I hereby declare that the Kerala University of Health and Allied Sciences,

Kerala shall have the rights to preserve, use and disseminate this dissertation in print or

electronic format for academic / research purpose.

Date : Signature of the Candidate:

Place : Name:
 ACKNOWLEDGEMENT

I hereby take immense pleasure to express my sincere thanks and gratitude to each and

every one without whose guidance and efforts, I could not complete this project work.

First of all let me thank The Almighty God for giving me this great opportunity.

The overall guidance and moral support for the completion of the project were given by

the staff of Mother Hospital.

Let me specially thank Mrs.Sajitha S .Nair, teaching staff of Mother College of

paramedical sciences, who supported me during every step of this project. She provided

all the technical support and various reference materials. She helped to correct the

mistakes and gave solutions to various problems which I faced during this study.

Once again I thank each and every soul especially my colleagues who supported me

during this project.

22-6-2016 DENNA FRANCIS

TABLE OF CONTENTS

Chapter Title Page No

1 Introduction

2 Objectives

3 Review of Literature

4 Methodology

5 Result

6 Discussion

7 Conclusion

8 Reference
ABSTRACT

BACKGROUND

The microorganisms are ubiquitous in nature and saprophytic pathogenic bacteria pose

risk of serious health problems. Computer keyboards and mice have been documented as

an environmental object or fomites serving as a source of microorganisms particularly in

the hospital setting. The wide spread use of computers in hospital set up, which are

handled by both patients and healthcare worker, the probability of it acting as an

inanimate vector is very high.

OBJECTIVE

The aim of this study was to assess bacterial isolates on computer keyboard and mice

and giving possible solutions about the same.

METHODOLOGY

A cross sectional study was conducted in Mother Hospital campus for the time period of

two months. Samples were collected from computers located in Mother Hospital by using

sterile cotton swabs. Then the collected samples were inoculated in to nutrient agar,

blood agar, Mac – Conkey agar, and incubated. The bacterial isolates were examined and

identified by colony morphology, Gram staining and biochemical characteristics.

RESULT

Growth was found in all samples. Among the total isolates 34 isolates were Gram

positive bacteria and 48 isolates were Gram negative bacteria. The isolates include

Bacillus species (50%), Staphylococcus aureus (6.6%), Klebsiella pneumonia (33.35),

Enterobacter (10%), Pseudomonas species (10%),Gram negative bacilli (25%),

Acenitobacter (3.3%).
CONCLUSION

The present study was conducted to isolate and identify pathogenic organisms on the

surface of computer. Computer keyboards and mice are the most open surface parts of

computer which showed 100% contamination. The bacteria identified have pathogenic

potential and hence their presence on computer surfaces may be additional reservoirs for

the transmission of microbes and become vectors for nosocomial infections in the

hospitals or healthcare setting and its surroundings. Hence the need for awareness on

cleaning of such surfaces or disinfection and adequate hand hygiene are of prime

importance.
INTRODUCTION
The use of computer and its associated accessories are becoming inevitable in a world of

various technological and informational transformations. Nowadays, computer has got

application in various fields of knowledge, work places and disciplines so much that, its

importance cannot be undermined because almost all of the operations are directly

dependent on computer usage. Owing to this indispensible nature of computer to the

various activities of man in this technologically dominated society, there is increasing

rate of interactions with computer from day to day.

As the popularity of computers increases, there is a need to recognize that the computer

equipment may act as a reservoir for the transmission of potentially hazardous or

pathogenic microorganisms. The computer can be act as fomites, especially computer

keyboard and mouse which are the most open surfaces of computer.

Some investigators have suggested that computer keyboards may serve as a reservoir for

some pathogens because of the increased use of computers in patient areas and contribute

to cross – transmission because of acquisition of transient hand carriage by healthcare

personnel during contact with the contaminated computer keyboard surface.

The intrinsic ability of some pathogens to survive in adverse environmental conditions

such as extreme temperature, pH, low oxygen pressure, and in the presence of little or no

nutrient and their ubiquitous nature inform researchers of the need to investigate

computer systems and especially the keyboards as media in infectious transmission. It has

been reported that computer may act as reservoirs for the transmission of potentially

hazardous or pathogenic microorganisms. These pathogenic microorganisms include

bacteria, fungi, protozoan, viruses, Chlamydia, and Rickettiziae. Each of these organisms

has developed one or more mechanisms of colonization and survival in any environment.

They have been reported to cause 98% of infections and diseases. Examples are

Staphylococcus aureus, Klebsiella, Pseudomonas, Escherichia coli, Listeria, and

Candida, Yesinia, Mycoplasma, Corynebacterium diphtheria etc.

Computer accessories in particular keyboards and mice are of high preference and have

been implicated in most cases as agents of infections transmission both in the community

and in the hospital environment. This is because they are the accessories with the highest

level of interaction.
OBJECTIVES

 To detect the surface contamination on computer keyboard and mouse.

 To suggest prophylactic measures for the same.

REVIEW OF LITERATURE

Computer has been described as the latest technological media

which are capable of receiving and accepting data, and performing

operation according to instructions (programme), and providing result of

the operation with great speed and accuracy[1] The importance of

computer had been identified in various sectors such as health,

agriculture, finance, education and research institutes[2] Its pace of

operation has made its application in these fields inevitable .The

inevitability of computer in most of the identified fields has been a

major factor for the continuous proliferation of computer usage in all of

these fields [3].

Computer technology has become a base necessity in the modern

everyday life. It has made most of our daily activities easier, faster and

more accurate from communicating to banking to even surgeries

.computers just like microbes are ubiquitous and they continue to have

an increased presence in almost every aspect of our occupational,

recreational and residential environment[4].

As the popularity of this facility increases, it has been discovered

that computer equipment may act as a reservoir for the transmission of

potentially hazardous or pathogenic microorganisms due to human and

environmental interactions[5].
 A search of literature has revealed that microorganism are

colonizing and contaminating environmental objects in home. In

hospitals, schools, day care environments, objects are contaminated by a

variety of pathogenic and non pathogenic microorganisms that can

persist on surface for prolonged periods [6].

The ability of a computer to act as a fomites has been previously

documented in healthcare (Huber and Pelon , 2005)and hospital

environment (Bures et al.,2000).The increased availability of multiple –

user computers in the hospital setting means that these items or

equipments are handled by numerous users on a daily basis[7].

Given that computers are not routinely disinfected, the opportunity

for the transmission of contaminating microorganisms is potentially

great[8].

Most people do not realize that microbes are found on many

common objects; outdoors in their offices and even in their homes.

Computer labs are another environment which is recently available in all

categories of scientific and technological departments [9].This can also be

a serious source of potentially hazardous or pathogenic microorganisms.

THE ROLE OF KEYBOARD AND MOUSE

The most important part of computer that has role in disease

transmission is keyboard and mouse. The oldest and the most widely

accepted keyboard layout is the “QWERTY keyboard “with the random

distribution of characters requiring more finger movements on the

keyboard[10].

These are touched by the bare hands of people who are in various

hygienic conditions. People believe that microbes are only present in

research laboratories or in hospitals and clinics and thus they have a

misleading feeling of security in other places. Lack of knowledge where

germs prowl could be the cause of health problems. Infact 80% of the

infections are spread through hand contact with hands or other objects

(Reynolds et al., 2005)[11].

This considered a serious issue; since patients are facing the risk of

contracting bacterial infections caused by these microorganisms can

spread to humans directly or indirectly via, inanimate objects called

fomites (keyboard and mouse).

The daily use of these computers by different users increases the

opportunity of microbe transmission and hospital acquired infections in

hospitals.

Health care - associated infections are an important cause of

morbidity and mortality in hospitals. Yearly more than 2 million patients

acquire health care –associated infections, resulting in 90000 deaths and

health care costs that are estimated to exceed $ 5 billion[12].

The computers keyboard and a mouse have a very dynamic

environment. Since the keyboard is constantly in contact with human

hands, there will be a stream of things leaving the hands and entering the

keyboard or of course, leaving the keyboard and entering the hands. In

the former case, bacteria from different niches could be transport over if

the hands have not been sanitized. In the later case, already pre-existing

bacteria will start to grow will transfer onto the hands and then on to the

new surfaces that the hand will come in to contact with. There are many

communities that come in to contact with the keyboard. In general, the

bacteria that lives on our skin, finger nails, hands, and anywhere the

hands have been are likely to transfer new bacteria over to the keyboard.

In a place where there is a lot of people moving in and out, such as a

hospital or office, there is likely to be a good number of people that are

sick, and through them comes the new bacteria that will eventually settle

on the keyboard through the air or from physical contact[13].

Computers have become a vital component of health care delivery

for improved and effective care. valuable patient related information is

available in computers at a click of button. Healthcare providers move

back and forth, between computers and patients while delivering

healthcare, as a part of daily routine. There are some reports on the

microorganisms colonizing computer keyboards in different locations of

hospital environment including clinical areas[14]. In a discussion of the

bacterial niche of a keyboard, so much depends on the user and what

bacteria and contaminants a user and what bacteria and contaminants a

user brings to the device.

 Significant amount of bacteria on keyboards in healthcare

environments is transferred through wet gloves or poor hygiene from

healthcare specialist. Bacterial transmission results from tapping on the

keys and regular usage of the device, which may incur contaminates

such as blood, secretions, or other various sticky substances in such a

health care environment (Fukata et al., 2008).From tests carried out,

95% of cultures from keyboards tested positive for microorganisms

though most were simple skin flora (Schultz, Maureen et al., 2003).The

focus of the research has been on pathogenic bacteria that pose threats to

nosocomial infections.

Having established from literatures that Computer Keyboards are

capable of hosting pathogenic microbes, and hence serve as portal of

infections, it is essential to identify the extent to which the public who

continually interact with Computer Keyboards are aware of the risk

associated with its possibilities as a portal of infection .To paucity of

literature on this issue necessitates the need to inform users about the

risk associated with this inevitable technological tools that is becoming

almost unavoidable in a developing societies[15]. So much as computer

system is a good technological tools, it also has the possibilities of

transferring microbial infections from one individual to another. Several

reports have revealed the possibilities of Computer Keyboards being a

medium for infection transfer. A recent report in the USA linked the

prevalence of nosocomial infections or cross contamination from the

hospital to the computers often used for diagnosis in the hospital. The

outcome of some studies in the United States revealed the isolation of

various species of bacteria, fungi and fungal spores[16]. Another study in

the USA revealed that Computer Keyboards contributes significantly to

the spread of hospital acquired infections, it was estimated to be

responsible for about twenty - five percent of microbes associated with

nosocomial infection(13) .Despite these findings the rate of interaction

with Computer Keyboards has remained on the high side because

Computer Keyboards has got applications in various economic sectors

.This thus increases the risk of contacting some of the infectious

organisms associated with Computer Keyboards surfaces[17]. The public

are exposed to this risk unconsciously because of the low level of

awareness among users of computers in all sectors, in particular

education and health sectors, this thus serve as a medium to inform users

of keyboards of the need to be more careful as they interact with this

technological gadgets[18]. There may be need to exchange computers

among users in a work place, cyber café as well as other public domain

this can serve as a means of transferring microbes as well as infections

amidst users.

Microorganisms on the skin are generally divided in to two

categories. Resident flora and transient flora. Resident flora is microbes

that normally colonize or live on the skin of most individuals; they

generally do not cause infections unless they are introduced to normally

sterile body sites and / or unless the host is immunocompromised. In

contrast, transient floras are microbes that are present on the skin for

only a short time. They tend to be more pathogenic than the resident

flora and are responsible for most nosocomial acquired infections[19]

These transient or contaminated floras may be picked up by the hands of

a healthcare worker; for example, when they touch a patient or any

contaminated objects, such as a computer component.

COMMONLY FOUND BACTERIAL AGENTS

The environmental conditions vary depending on temperatures

around the keyboard and whether or not the keyboard is on a laptop[20].

If the keyboard is on a lap top it could possibly provide heat and

moisture for long enough durations to have an effect on bacteria such as

Enterococcus, which is known to survive a wide range of environmental

conditions. Studies have shown, however, that the presence of serum or

albumin (known contaminants on keyboards touched by wet gloves),

and a low temperature, with high humidity results in longer life time of

bacteria on contaminated surface. Many nosocomial pathogens can also

survive on dry inanimate surfaces for months (Kramer et al., 2006).

Staphylococcus aureus usually found on skin or in the nasal

environment and only survives on dry skin on the outside of the body.

Appears on keyboards quite a bit as a result of usage. Methcillin

resistant strain of Staphylococcus aureus found on keyboards a high

percentage of the time in hospital environments which can cause

infections in patients (Fukata et al., 2008).Clostridium perfringens

usually found in human gastro intestinal tracts and environment such as

sewage and soil, however, in a healthcare environment can cause gas

gangrene. Probably will not survive long on a keyboard as its primary

target is living tissue. Found on keyboards at lesser degrees.

Enterococcus bacteria are usually find in the bowl and are known

to be able to survive adverse conditions that other bacteria usually won’t

grow in. They are known to survive at temperatures of 60c and in

anaerobic conditions with varying degrees of acidity. Enterococcus

species represents some of the highest rates of appearances in hospital

environment keyboards (Hartman, et al., 2004). Corynebacterium

commonly found on human skin and mucus membranes. Streptococcus

mostly anaerobic while some are facultative anaerobes, they do not carry

out oxidative phosphorylation and can survive in more acidic

environments which might be present on keyboards. Medically

significant Streptococci require lots of amino acids, vitamins, and

nutrients thus are not normally found isolated in an environment [21]

Many bacteria have been isolated from computer keyboards and

mice in worldwide including developed countries.

Bacteria pathogens commonly found from computer keyboards

and mice that can able to survive for long period of time and resist

disinfection are important for computer keyboard and mice

contamination. In different literature from nosocomial pathogens

indicated that Gram positive bacteria, such as Staphylococcus aureus,

Enterococcus species and Streptococcus pyogenes survive for months on

computer keyboards and mice[22]. Many Gram negative bacteria, such

Escherichia coli, Klebsiella species, Acenitobacter species and

Pseudomonas aeroginosa can survive on computer keyboards and mice

surfaces even for months. The degree to which computer keyboards and

mice are contaminated is different. One study reported, for example

studies from 100 keyboards in 29 clinical areas, 95% keyboards were

[23]
positive for microorganisms . Based on the level of pathogen on

keyboards 25% of keyboards in hospitals carry pathogens at any given

time more than double that of other commonly-touched surfaces [24].

The ability for computers to act as fomites has been documented in

hospital and health care environments and contamination of the

environment including the computer keyboard with bacteria is nowadays

have got special recognition in various parts of the world[25].

Some investigators have suggested that computer keyboards may

serve as a reservoir for some pathogens because of the increased use of

computers in patient areas and contribute to cross contamination because

of acquisition of transient hand carriage by healthcare personnel during

[26]
contact with the contaminated computer keyboard surface . The
concern has been raised that contact with contaminated computer

keyboards might serves as a mechanism for contaminating the hand of

health care workers with potential pathogens, thereby leading to cross

[27].
contamination of patients Even though the role of hospital

environments as a reservoir of nosocomial pathogens is controversial,

the introduction of bedside computers in to patient’s room in the critical

care environment may play a role in the transmission of nosocomial

[28].
pathogens Undisputedly hands are the main source of pathogen

transmission. Cross contamination of microorganisms by the hands of

care personnel from computer components at the patient’s bedside,

might introduce an additional risk for critically ill patients considering

the frequent contact of nursing and medical staff during patient care with

these fomites [29].

Keyboard use has been on the increase and the high rate of

unhygienic interaction with the computer made them a potential

reservoir of microbial pathogen [30]. Microorganisms are ubiquitous and

have devised various means of survival in any environment and surfaces,

including some that do not even provide them with the adequate nutrient

for their growth .This ability in microorganisms enabled them to

colonize and remain on the surface of computers and act as fomites.

Several microorganisms have been isolated from Computer Keyboards

surfaces [31]. One characteristic feature of all these microbes is that they

have peculiar mechanisms in surviving in harsh environmental

conditions; such as encysting, encapsulation and spore production .Most

of the microorganisms that have been isolated from the Computer

Keyboards have been linked with different health related conditions

particularly in nosocomial infections. Apart from being associated with

hospital acquired infections, this organisms that have been isolated from

computers are also common in the community, hence the need for

cautions when exchanging and interacting with computer systems. The

common associated organisms include; bacteria(Bacillus species

,coagulase negative staphylococcus aureus , oxacillin sensitive

Staphylococcus aureus ,oxacillin resistant staphylococcus aureus

,vancomycin sensitive enterococcus , vancomycin resistant enterococcus

, diphtheroids ,Micrococcus species ,Bacillus species ,non fermentative

gram negative rod Propionibacteria , Pneumococci ,Viridans

streptococci), fungi and fungal spores. The fungi species that have been

linked to Computer Keyboards are mainly Aspergillus species .These

microorganisms are associated with various infections that effect human

health both systemically and locally [32].

THE COMMON HOSPITAL - ACQUIRED INFECTIONS

1. Aspergillosis

2. Respiratory tract infections

3. Pneumonia

4. Dysentery

5. Cholera
 These thus pose Computer Keyboards as potential agent for the

transmission of infection.

Not only in the health sector, as the computer got application, but

also in various other sectors of the economy. This suggests the

possibilities of the public contacting some of the infectious diseases that

had been with computer keyboard.

Fomites or inanimate objects have been identified with the

potential characteristics of serving as a reservoir for microorganisms and

their spores hence the possibility of contacting these microbes or their

spores through the use of these agents. Computer is a form of inanimate

object that have also been identified as a reservoir that host microbes or

microbial spores [33].

Computer Keyboards and mouse are central to computer as a reservoir

of pathogenic microorganisms capable of causing infections in users

with capacity to transmit to other users or non users in the community

through interaction. In Australia, pathogenic microorganisms such as

bacteria, protozoa cysts and even fungi, their spores and bacteria spores

have been identified on computer.

It has long been recognized that inanimate objects in the patient’s

environment can harbor microorganism .These objects might be medical

[34]
tools, such as stethoscopes ear thermometers, or bronchoscopes, or

common non medical objects, such as ball point pens, bedrails and

beside tables, and especially computer keyboard and mouse. Only

recently have investigators begun to examine the microbial

contamination on computer hard ware and to ask if these

microorganisms might play a role in patient acquired infections.

FACTORS INFLUENCE THE LINK BETWEEN

COMPUTERS AND PATIENTS

Two factors that play a role in the link between any fomite, such as

a piece of computer hard ware, and the patient are the ability of a

particular microbe to survive on a particular surface and the fact that

various vectors, such as health care workers, can transfer

microorganisms from one surface to another. Microorganisms survive

for different periods of time on different surface. Survival varies

depending upon the particular microbe, the particular surface, and the

concentration of the microorganism on the surface.

In general, the greater the concentration of the microbe, the longer

it survives. Survival can range from minutes to months. Obviously, if a

microbe only survives for a few minutes on an inanimate object, such as

the computer terminal, then the possibility of that microbe being

acquired by a patient is quite small .However, conversely, if particular

bacteria or fungi survive for weeks to months on a certain surface, then

the odds of that organism being picked up by a patient or health care

worker are considerably increased.

 PLASTIC COVER THREATS

Most of the accessible components of computers are made of

plastic. In a series of studies in which the survival of a variety of bacteria

and fungi where determined on a number of different fabrics and plastics

(including the plastic skins used to protect computer keyboard),

microbial survival was often days to weeks on both types of surfaces

.However, when there was a difference in survival between the fabrics

and the plastics, the microbes tended to live longer on plastics. Hence,

the long survival times of certain microorganisms, particularly on

plastics such as those associated with computers, contributes to the

possibility of computers acting as reservoirs for these microbes [35].

Cleaning our keyboard and mice regularly is a smart solution that

most people ignore; only about half of computer users in the average

office environment clean their computer keyboards at least once a

month. Of course, sharing a keyboard with multiple people makes it a

much more dangerous surface when it comes to passing diseases. If we

use our own keyboard and mouse and nobody else uses it, then the

chances of that keyboard and mouse serving as a method of transmission

is fairly small, but where sharing keyboards is concerned there is a

higher probability of transmission occurring. We can clean in between

the keys of our keyboard with compressed air (i.e. canned air ) on a

monthly basis, and we can also use canned air to blow out the fans and

vents on our computer regularly too. We can also wipe the surface of the
keys and keyboard with antibacterial wipes regularly for the best

protection.

CLEANING AND DISINFECTION

Cleaning is the removal of all foreign material, such as dirt and

organic material from an object .Sterilization is the complete elimination

or destruction of all forms of microbial life, while disinfection is a

process that eliminates many or all pathogenic microorganisms, with the

exception of bacterial spores, from inanimate objects. Since there are

relatively few situations in which computer hardware would need to be

disinfected. Because dirt can harbor microbes from the normal

disinfecting process, successful disinfection should be preceded by

cleaning. However, certain disinfectant phenols may accomplish both

tasks in one process.

There is no perfect disinfecting agent: each chemical has its own

advantage and disadvantage, depending on the situation in which it is

used. Therefore, in any medical facility, the infection control personnel

should be consulted about appropriate cleaning or disinfecting agents

and procedures, Factors to be considered include the level of disinfection

necessary for that particular computer, the potential types of organic and

microbial contaminations that might be present, and the cleaning or

disinfecting agents available.

 For disinfection process, one needs to assess the compatibility of

the disinfecting chemical with the computer hardware to be cleaned or

disinfected. Many chemical disinfectants required that the surface to be

disinfected be exposed to the liquid disinfectant for 10 minutes. Such

exposure could create an electrical or corrosive problem to certain pieces

of computer hardware.

In some circumstances, such as the computer keyboard, the

problem of chemical damage to the keyboard components can be

elevated by the use of a thin plastic keyboard cover (aka skin), which

can be liberally soaked with disinfectant without fear of compromising

the computer.
MATERIALS AND METHODS

STUDY AREA AND DESIGN

The study was conducted in the microbiology department of Mother Hospital, Thrissur,

Kerala, during a period of May 2 to June 30

The research was focused on the microbial studies of pathogenic bacteria on computer’s

mice and keyboards collected from different computers of Mother hospital campus.

A total of 60 samples were collected (30 from computer keyboards and 30 from mice).

The computers were in use by both clinical and non clinical staff including doctors,

laboratory technologists, nurses, recording officers, and personal computers from

different health care works, all of whom had direct or indirect patient contact.

The areas from which samples have been collected are:

1. Laboratory room

2. Recording room

3. Reception

4. Clinical staff office room

5. Personal computers of heath care workers.

REQUIREMENTS

1. Cotton swab sterilized (160˚c for 1 hour in hot air oven)

2. Test tubes

3. Incubator

4Inoculating wire loop

4. Bunsen burner
MEDIA USED

1. Peptone water

2. Plating media ( Mac – conkey agar , Blood agar )

3. Biochemical media ;

a) Christensen’s urease agar medium

b) Simmons’s citrate medium

c) Glucose phosphate broth for methyl red reaction

d) Glucose phosphate broth for Vogues - Prauskuer reaction

e) Mannitol motility medium

f) Triple sugar iron agar

g) Peptone water for Indole reaction

h) Hugh – Leifson medium for oxidation – fermentation test

REAGENTS USED

1. Hydrogen peroxide (3%)

2. Tetra- methyl- paraphenylene- diaminedihydrochloride (1%)

3. Human EDTA plasma

4. Kovac’s reagent

5. Methyl red reagent

6. Barret’s reagent

7. Paraffin oil (heavy)

METHOD OF SAMPLE COLLECTION

Isolation of various bacterial contaminants from these two objects (computer keyboard

and mouse) was performed through standard technique.

Samples were aseptically collected from each computer keyboard and mouse by using

sterile moistened cotton swabs before cleaning and disinfecting computer keyboards and

mice with damp cloth and alcohol (70% methylated spirit wipes).

The swab moistened with peptone water is used for sample collection. The moistened

swabs was rotated over all the keys (letter keys, space bars, enter keys, function keys,

number keys, and other keys) and from palm rest left and right click buttons of the mouse

were aseptically collected.

While swabbing on surface different shapes, appropriate care was taken to cover

approximately the same surface area.

The collected swabs were put in to separate sterile tubes, which contain 5 ml peptone

water. Swabs were immediately taken to microbiology laboratory.

DAY – 1

On reaching laboratory, the tubes are thoroughly vortexed for 1 minute. In the

same manner all the 60 samples of dust containing sterile cotton swabs were dipped

separately in to the peptone water and were left in incubator for overnight at 37˚c.

DAY – 2

After 18 – 24 hours of incubation, the peptone water became turbid; turbidity

ensured the presence of certain bacteria in the sample. For identification of bacterial

colonies; the sample suspension is inoculated to media like;

Nutrient agar: For morphological identification of bacteria

Blood agar: For the growth of heterotrophic bacteria

Mac – conkey agar: For identification of coliforms.

The above mentioned agar plates were prepared aseptically small amount of culture was

taken with the help of inoculating loop from the peptone water medium and were

streaked on to the agar plates.

The agar plates were incubated at 37˚c for 18 – 24 hours. All the plates were incubated

aerobically.

DAY – 3

After 24 hours of incubation, the plates were examined. Bacterial colonies

differing in size, shape, and colour were selected from the different plates and further sub

cultured on to peptone water.

The pure colonies from which further identification were done by using standard

microbiological techniques.

1. Colony morphology

2. Preliminary biochemical tests

3. Gram staining

4. Motility testing

5. Biochemical tests

COLONY MORPHOLOGY

The colony morphology includes;

In blood agar, based on type of haemolysis the bacteria can be classified. The organism

shows alpha-haemolysis (partial haemolysis with a green hallow), Beta – haemolysis

(complete haemolysis), gamma haemolysis (no haemolysis)[44] .

Mac – conkey agar is a differential and low selective medium used to distinguish lactose

fermenting (pink colour), and non lactose fermenting (colorless)

PRILIMINARY BIOCHEMICAL TESTS

The suspected colonies are separately inoculated into 5ml peptone water and

incubated at 370c for 4 hours. these are used for large and quick yield of suspected

bacteria. The suspected colonies are subjected to preliminary biochemical tests.

1. Catalase test

2. Oxidase test.

3. Coagulase test.

CATALASE TEST.

This demonstrates the presence of catalase, an enzyme that catalyses the release

of oxygen from hydrogen peroxide. The catalase test should be performed from a

medium that does not contain blood, since RBC themselves may produce a weakly

positive catalase reaction. Colonies also transfer with the help of platinum loop or

wooden stick.

The catalase test is performed by emulsifying bacterial colonies with 3%hydrogen

peroxide on a glass slide. Immediate and vigorous bubbling indicates conversion of the

hydrogen peroxide into water and nascent oxygen in the presence of catalase enzyme

produced by the bacteria.

OXIDASE TEST.

This test depends on the presence of certain oxidase enzyme present in bacteria

that will catalyses the transport of electrons between electron donors in the bacteria and a

redox dye, tetra- methyl-para-phenylene-diaminedihydrochloride.The dye is reduced to a

deep purple color.

A colony from the growth medium is removed with an applicator stick and rubbed on to

the disc (impregnated with the reagent).thee development of a blue purple color within

30seconds is a positive test. No color development within this time is a negative test.

COAGULASE TEST.

It is most commonly used biochemical reaction for the identification of

staphylococcus aureus.coagulase test differentiates staphylococcus aureus from CONS.

Staphylococcus aureus produce 2 forms of coagulase; bound and free coagulase.

BOUND COAGULASE TEST (SLIDE TEST)

Bound coagulase or clumping factor is bound to the bacterial cell wall and react

directly with fibrinogen. This results in the alternation of fibrinogen so that it precipitate

on bacterial cell causing the cell to clump when a bacterial suspension is mixed with

plasma.bound coagulase can be detected by slide coagulase test.

The suspected colonies are emulsified with a drop of rabbit EDTA plasma on a clean

glass slide. Macroscopic clumping in 10 seconds or less than 10 seconds indicates

positive reaction. No clumping indicates negative reaction.

FREE COAGULASE TEST (TUBE TEST).

Free coagulase is an extra cellular protein enzyme that causes the formation of a

clot when staphylococcus aureus colonies are incubated with plasma. The clotting

mechanism involves activation of plasma coagulase reacting factor (CRF) which is a

modified thrombin molecule to form a coagulase -CRF complex. This complex in turn

reacts with fibrinogen to produce a fibrin clot.

0.1ml of an overnight broth culture or an agar culture suspension of the organism is

mixed with 0.5 ml of a 1 in 5 dilution of human or rabbit plasma. In case of a positive test

the plasma clots and does not flows when the tube is inverted. No clot indicate negative

test.

GRAM STAINING

It is the most commonly used staining method derived by the histologist Hans

Christian Gram (1884) to stain bacteria in tissue sections.

It is the differential staining method, which differentiate organisms into gram

positive and gram negative according to their gram reaction. the gram stain is used to

categorize bacteria on the basis of their forms; size, cellular morphology, and gram

reaction.however,in a clinical microbiology laboratory ,it is a critical test for the rapid

diagnosis of infectious agents and help in assessing the quality of clinical specimens.

Gram stain makes separation of all bacteria into two broad classes; gram positive

bacteria are those that resist decolourisation and retain the primary stain, appearing violet,

gram negative bacteria are decolorized by organic solvents and therefore, take the counter

stain, appearing red.

 1. Prepare a thin smear and fix it by flaming.

2. Primary staining; cover smear with pararosaniline dye such as crystal violet,

methyl violet ,or gentian violet, and allow to react for 60 seconds.

3. Pour off primary stain and wash with water.

4. Apply grams iodine for 60 seconds.

5. Pour off iodine and wash with water.

6. Decolorize the smear with an organic solvent; absolute alcohol, acetone or aniline

for 1-2 seconds or until the colour oozes from the slide.

7. Wash with water.

8. Counter stain with a dye of contrasting color;saffranine,carbol fuschin or neutral

red for 60 seconds.

9. Wash with water and blot dry.

10. Observe the slide under 100 xs after putting a drop of cedar wood oil.

MOTILITY TEST

HANGING DROP METHOD:

It helps to distinguish motile bacteria from non motile bacteria. A drop of liquid

culture is placed on cover slip is inverted over a cavity slide so that drop remains

hanging. This prepation is then observed under low power to adjust edge of the drop and

under high power to study the motility of organisms.

BIOCHEMICAL MEDIA

.4 hour pure culture of each organisms are inoculated in to biochemical media and

incubated at 370 c for 18 – 24 hours.

INDOLE TEST

Indole, a benzyl pyrole is one of the metabolic degradation product of amino acid

tryptophan. Bacteria that possess tryptophanase are capable of hydrolyzing and

deaminating tryptophan with the production of indole, pyruvic acid and ammonia.Indole

is then test for by a colourimetric reaction with paradimethylaminobenzaldehyde.

In to 48 hour culture suspension add 0.5 ml Kovac’s reagent and shake gently. A red

colour in the alcohol layer indicates a positive reaction.

METHYL RED TEST

This test is used to detect the production of sufficient acids during the

fermentation of glucose and maintenance of the ph off an old culture below pH 4.5 which

can be detected by change in the colour of methyl red indicator which is added at the end

of incubation. The indicator is red at ph 4.5 which indicate a positive result, yellow at ph

6 indicate negative result. The media used is glucose phosphate broth.

V- P TEST

This is the test used to determine the ability of some organism to produce non

acid products like acetyl methyl carbinol (acetoin ) from glucose fermentation. In the

presence of alkali and atmospheric oxygen, acetoin is oxidized to diacetyl which reacts

with α- naphthol in reagent (Barrett’s) to give red colour. The media used is glucose

phosphate broth.
CITRATE UTILIZATION TEST

The test is used to detect the ability of an organism to utilize citrate as the source

of carbon and energy source for its growth; with resulting alkalinity. This change in ph

indicated by the blue colour of the medium. The medium used is Simmons’s citrate agar.

UREASE TEST

This test is done to determine the ability of an organism to produce an enzyme

urease which splits urea to ammonia. Ammonia makes the medium alkaline and thus

phenol red indicator changes to pink or red in colour.The medium used is Christensen’s

urease agar.

OF TEST

This test is used to determine whether substrate utilization is oxidative or

fermentative process. Oxidative process requires oxygen whereas fermentative do

not.utilisation detect by observing whether acid byproducts are formed in presence or

absence of oxygen. Production of acids in the medium can be detected by noting the color

change of the ph indicator incorporated in the medium.

TRIPLE SUGAR IRON AGAR TEST

TSI contain three sugars; glucose (0.1%), lactose (1%), sucrose (1%).Ferric

ammonium citrate and phenol red as indicator. It contains a slant and butt. The slant

portion is exposed to atmospheric oxygen, is aerobic. The lower portion called butt is

protected from the air and is relatively anaerobic.

If only glucose is fermented after 24 hours of incubation an alkaline slant (red0 and acid

butt (yellow) is observed . The production of hydrogen sulphide is manifested as a black

precipitate in the medium, which is due to the production of ferrous sulphide. Gas
production can manifested as single or multiple bubbles, splitting the medium or pushing

up the entire plug of the medium. The organisms are inoculated by first stabbing in to the

butt, withdraw through the same line and stroke over the slant by using charged

inoculating straight wire.

Yellow/yellow (initial reaction after 12 Only glucose is fermented so acid slant

hours) and acid butt initiate with or without

gas production.

Yelloww/yellow (after 24 hours ) The organism frmentt glucose +

succrosse or glucose+lactose and all

three without gas production

Red/yellow later the organism starts utilizing

peptone results in alkaline ph on the

slope and acid butt results with or

without gas

Red/red No fermentation of glucose, sucrose

and lactose, there is no change in the

medium.

MANNITOL MOTILITY TEST

If the organism is mannitol fermenting there will be colour change from red to

yellow. If the organism is motile it produces diffused growth that spread throughout the
medium, making it slightly opaque. Non motile organism does not show any diffused

growth, it only grows along the stabbed line.

DNASE TEST

DNAse is an enzyme that hydrolyzes DNA. Production of DNAse can be detected

by growing organism in a medium containing DNA. The medium is pale green because

of DNA methyl green complex. Around the colonies of DNAse producing organism

DNA will be hydrolyzed. Addition of HCl results in the precipitation of unhydrolysed

DNA ,this makes the medium opaque and clear zone around DNAse producing organism.

.
RESULT

I conducted a prospective study to question the presence of pathogenic bacteria on

the computer keyboard and mice surface in and around a hospital environment and its

importance in hospital acquired infections in patients.

The main objective of the present microbial study was to isolate and identify the

pathogenic microorganisms on the external surface of computer’s mice and keyboards to

create public awareness about the health hazards resulting from these pathogenic

microorganisms. To conduct this study, total 60 samples were collected from different

computers used in Mother Hospital located in Thrissur, Kerala. The computers were used

by both clinical and non clinical staffs who have direct or indirect contact with patients.

A simple random technique was used to sample 30 keyboards and mice from hospital

surroundings by swabbing their surfaces. All the samples were first cultured in peptone

water, and nutrient agar, which ensured the presence of certain pathogenic

microorganisms. For further conformation and identification, the culture from the nutrient

agar was streaked in blood agar and Mac – conkey agar media. The presence of

pathogenic bacteria like Bacillus species, Staphylococcus aureus, Klebsiella pneumonia,

Enterobacter species, Acenitobacter species, Pseudomonas species, Gram negative

bacilli were confirmed on the basis of Gram staining, colony morphology, Motility

testing, and biochemical testing.

A list of pathogenic microorganisms has been identified to be associated with computer

keyboards and mice.

BACTERIA TOTAL

PERCENTAGE

Bacillus species 50%

Staphylococcus aureus 6.6%

Klebsiella pneumoniae 33.3%

Enterobacter species 10 %

Pseudomonas species 10%

Gram negative non 25%

fermenting bacilli

Acenitobacter 1.6%

Table; 1Bacterial species isolated from the surface of computer keyboards in percentage.

Diagram1; Distribuition of bacterial isolates on computer surface.

10% Bacillus
10%
10% staph.
50%
Kleb.
10%
Enteroba
33.30% Psedudo.
GNB
6.60%
Acenito.
 HOSPITAL COMPUTER ISOLATES
BACTERIA
Keyboards Mice Total

Number percentage Number Percentage Number Percentage

Bacillus species 16 53.3% 14 46.6% 30 50%

Staphylococcus aureus 3 105 1 3.3% 4 6.65

Total Gram positive bacteria 19 44.18% 15 38.46% 34 41.46%

Klebsiella pneumoniae 10 33.3% 10 33.3% 20 33.3%

Enterobacter species 3 10% 3 10% 6 10%

Pseudomonas species 2 6.6% 4 13.3% 6 10%

Gram negative bacilli 8 26.6% 7 23.3% 15 25%

Acenotobacter 1 3.3% 0 0% 1 3.3%

Total Gram negative bacteria 24 55.18% 24 55.18% 48 55.18%

Table:2

A total of 60 (30 from keyboards and 30 from mice)samples were collected. All the

samples collected yielded growth; however the extent of contamination varied. (Table; 1)

From the total bacteria isolates (82), 34 (41.46%) were gram positive bacteria and

48(58.53%) were gram negative bacteria. Majority of the samples collected from

computer keyboard and mice had multiple bacterial growth.

Among Gram positive growth Bacillus species were predominant ie; 50%.

Staphylococcus aureus(6.6%) was also isolated. A total of 48 Gram negative bacteria

isolated of which Klebsiella pneumonia (33.3%) were predominant. Other Gram negative

bacteria include Enterobacter(10%),Pseudomonas species(10%), Gram negative

bacilli(25%), and Acenitobacter (1.6%)

Bacteria Ind. MR VP Cit. Ure. Cat. Oxi. Coa. OF TSI Mann.

Bacillus species

Staphylococcus - + + + - + - + Fer. A/A +

aureus
Klebsiella pneumonia - - + + + + - - Fer. A/A +

Gram negative bacilli - - - - - - - - Ass. K/NC -

Enterobacter species - - + + + + - - Fer. A/A +

Pseudomonas species - - - + + + + - Oxi. K/NC +

Acenitobacter species - - - - - - - - Ass. K/NC -

Ind.(Indole), MR(Methly red), VP(Vogus- Prauskauer), Citrate(Citrate), Ure(Urease),

Cat.(Catalase), Oxi.(Oxidase), Coa.(Coagulase), OF(Oxidative – Fermentation),
TSI(Triple Sugar Iron), Mann.(Mannitol).

Ind.(Indole), MR(Methly red), VP(Vogus- Prauskauer), Citrate(Citrate), Ure(Urease),

Cat.(Catalase), Oxi.(Oxidase), Coa.(Coagulase), OF(Oxidative – Fermentation),
TSI(Triple Sugar Iron), Mann.(Mannitol).

20

10
Keyboard
mouse
0
DISCUSSION

Computer use has been on the increase and the high rate of unhygienic

interactions with the computer made them a potential reservoir of microbial pathogen.

This ability of microorganisms enabled them to colonize and remain on the surface of

computers and act as fomites.

Most of the microorganisms that have been isolated from the computer keyboard have

been linked with different health related conditions particularly in nosocomial infections.

The common associated organisms include Bacillus spps., Klebsiella pneumonia,

Enterobacter spps., Pseudomonas spps., Acenitobacter spps., Non fermenting Gram

negative bacilli[36].

In this study, all the samples collected yielded growth (100%). However the extent of

contamination varied. In the present study Gram negative isolates were more prevalent

than gram positive bacteria. Multiple bacterial isolates were reported from almost all

samples. The Bacillus species were most abundant (50%) which being a normal

contaminant in atmospheric air.

The study reveals 6.6% prevalence of Staphylococcus aureus species (3 from keyboard

and 1 from mouse).This may be contamination from the skin, mouth or nose of the

computer handlers and soil which might be introduced directly into the computers was

responsible for the colonization of computers. Previous studies have reported that

bacterial contamination occur computers surface s located in a college setting and may

reflect the multiple user environment where the possibility of contamination by

individuals who are carriers of bacteria such as Staphylococcus aureus is greater and the

isolation of viable microorganisms suggest that the species present are able to persist for
a period of time on these surfaces [37]. As with hospital settings, computer keyboards and

mice in tertiary institutions may act as a vehicle for the transmission of these organisms.

In the present study, gram negative bacteria were more in number. The Klebsiella

pneumonia were the second dominant pathogen with over all isolation rate of 33.3% (10

from keyboard and 10 from mouse) The isolation rate of other Gram negative isolates

include ; Gram negative bacilli (25%), Pseudomonas species(10%), Enterobacter species

(10%), Acenitobacter species (3.3%)

A brief report by Isaacs et al., gives a similar result to that of present study. 27 keyboards

in a burn unit were swabbed from which bacteria like Staphylococcus aureus and

Pseudomonas, both of which are capable of causing serious infections in burn patients,

were found.

Multiple studies shown that computer keyboards could harbor several colonies of

bacteria; for instance a study conducted at the Henry Ford Hospital in Detroit showed that

keyboards located in triage and registration areas were more contaminated with bacteria

than those in other areas of the emergency department. In their study 72 standards, non –

silicone rubber keyboards were swabbed on two different days, six days apart. All

keyboard key, except the function keys, were cultured and analyzed for bacteria. The

results showed that less than 14%, or 10 keyboards, were colonized with nine different

bacteria. Of the keyboards in non- treatment areas, nearly 32% were contaminated, versus

less than 9% in treatment [38].

This is in close connection with the present study which suggested that computer

keyboards may contribuite to cross - transmission because of acquisition of transient hand

carriage by healthcare personnel during contact with the contaminated computer

keyboard surface.

Forbes reports that harmful bacteria can linger on computer keyboards in hospitals,

making it easy for the germs to spread to patients [39].

Schultz et al., also demonstrated [41] that microbial contamination of computer keyboards

was prevalent and that commensals skin organisms were the most common

contaminating microbes. It was also reported that the degree of contamination was high

enough to potentially allow transmission via contaminated hands. The present study also

shows the commensal prevalence on computer surfaces [43].

Different studies in various parts of the world had assessed the extents of bacterial

contamination of computer keyboards and mice. For example, a study in USA the

isolated bacteria were Staphylococcus aureus (4%) and Enterococcus (12%)[40] This

results are almost similar to present study which have Staphylococcus and Enterococcus

prevalence of 6.6% and 10% respectively. The non fermenting Gram negative rod shows

higher prevalence (36%) than present study.

CONCLUSION

The present study was conducted to isolate and identify pathogenic organisms on the

surface of computer. Computer keyboards and mice are the most open surface parts of

computer which showed 100% contamination. The bacteria identified have pathogenic

potential and hence their presence on computer surfaces may be additional reservoirs for

the transmission of microbes and become vectors for nosocomial infections in the

hospitals or healthcare setting and its surroundings.

Hence the need for awareness on cleaning of such surfaces or disinfection and adequate

hand hygiene are of prime importance.

The possible solutions include;

 Replacement of traditional keyboards with silicone rubber models.

 Use of infection resistant keyboards which are completely flat and covered with an

hypoallergenic material resistant to bug growth.

 Use of keyboards-timed warning light which can be configured to meet the needs of

different environments. The warning lights would go out when all the sensors are

activated, indicating that the keyboards are clean[42].

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