INTERNATIONAL JOURNAL OF SCIENCE SCHOLARS, Vol-1, issue-03, pp.

177-199, 2017
Advance access publication- 17 August, 2017
This paper is available online free of all access charges

ISSN NO- 2456-8929

INTERNATIONAL JOURNAL OF SCIENCE SCHOLARS

The House of the Written Word

RESEARCH PAPER

ISOLATION AND IDENTIFICATION OF MICROORGANISMS FROM MOBILE

PHONES IN COLLEGE ENVIRONMENT

Jaspreet Kaur1*

1. Post graduate government college for girls, Department of Biotechnology, Sector 42,Chandigarh

Received- 20 July 2017; Revised 4 August 2017; Accepted- 17 August 2017

Abstract
The aim of this study is isolation and identification of bacterial colonization on mobile
phones. 3 samples were collected from the Teaching, Non-teaching and Student of the
PGGCG, sector-42, Chandigarh. For each mobile phone, sterile swab moistened with normal
saline was rotated over the surface of both sides of the mobile phone. For each mobile phone,
sterile swab moistened with normal saline was rotated over the surface of both sides of the
mobile phone. Swabs were cultured on Mac-conkey agar, nutrient agar, Sheep Blood agar,
Mullor hinton agar for bacterial growth. Plates were incubated aerobically at 370C for 48h
for bacterial growth. Positive isolates were identified according to the standard
microbiological techniques. All bacterial isolates were tested by available antibiotic disks.
The organisms consistently isolated in this research, with their frequency of occurrence were
Staphylococcus species (24.4%), E.coli (15.5%), Streptococcus species (13.3%), Bacillus
species (13.3%), Citrobacter species (8.9%), Enterobacter species (8.9%), Pseudomonas
species (6.7%), Serratia species (6.7%) and Pneumococcus (2.3%). The results therefore
highlight the health implications of using other peoples phones as they could be loaded with

177 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

microorganisms capable of causing various diseases and infections. Thus, cell phones could
serve as vehicles of diseases.
Keywords- Bacteria, Mobile phones, Diseases, Antibiotic disks.

INTRODUCTION-

With recent advances in the sources of communication, use of mobile phones has become
essential in every moment of life. A mobile phone (also known as a cellular phone, cell phone
or a hand phone) is a device that can make and receive telephone calls over a radio link whilst
moving around a wide geographic area. It does so by connecting to a cellular network
provided by a mobile phone operator, allowing access to the public telephone network. By
contrast, a cordless telephone is used only within the short range of a single, private base.
Mobile phone is a widely used personal gadget essential in daily life and is usually kept in
close contact with the body. It is used for communication by different group of people at
every place or situation including laboratory, lectures hall, auditorium and in class rooms.
Today, mobile phones have become one of the most indispensable accessories of professional
and social life. Although they are usually stored in bags or pockets, mobile phones are
handled frequently and held to the face [1, 2].
A mobile phone is a long range, portable electronic device for personal telecommunication.
With the invention of fully loaded mobile phones, in addition to the standard voice function,
a mobile phone can support many additional services such as SMS, email, pocket switching
for access to the Internet, and MMS for sending and receiving photos and videos [3].
Bluetooth for exchanging important data files, images. Latest applications include monitoring
security of house via online access of house for 24 hours with locking facility from far
distances. Keeping an eye on important keep up of personal details, files etc. Automatic
dialing of emergency numbers and messaging for help and telling them exact location for
help and immediate attention are now a days common. Connecting of mobile with Luxury
cars and automated transfer of calls on to the audio system installed on the cars. The
availability of prepaid services, where the subscriber does not have to commit to a long term
contract, has helped fuel for growth of cellular subscribers on a massive scale in India. With
high level of mobile phone penetration, a mobile culture has evolved, where the phone
becomes a key social tool and people rely on their mobile phone address book to keep in
touch with their family and friends. These devices not only help individuals share information
with each other, they are increasingly being used to help individuals gather information about
themselves [4].

178 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

Smart phones mobile phones with built in applications and internet access have rapidly
become one of the worlds most sophisticated self-tracking tools. Mobile phones serve as
clocks, organizers, reminders, calculators etc., depending on the mobile phone accessories. It
is considered that modern life is almost impossible to imagine without the use of mobile
phones. With all the achievements and benefits of the mobile phone, it is easy to overlook the
health hazards arising out of the excessive and unusual use of this item. Research has shown
that mobile phone could constitute major health hazard [3].
The electromagnetic radiation emitted from phones and base stations has a threat to lives
because, the electromagnetic radiation has been reported to alter the electric activity of the
brain causing sleeplessness, headache, malaises, memory retentiveness and low sperm
quality. It damages the DNA of manufacturing sperm cell.The excessive use of mobile
phones in general on environmental issues states disappearance of common sparrow from
urban areas of the country. Also individually if kept near heart tends to be risky for cardiac
health and if kept near gonads can affect reproductivity or normal functioning of gonads of an
individual. Excessive use of ear plugs for listening music can create problems related to
listening and dumbness.
Latest technology of android based mobiles dual blow(microbial and personal information
like habits and other personal details which one never would like to expose it to an unknown
person anywhere in the world) Forgetting the mobile at any place extra duty in ones
personal life .Compelled attention seeker a mobile phone. Negative aspects for any
particular pathogenesis linked with the mobile phone Radiations emitting from mobile
towers. Scope for survival of capsular species of bacillus and cocci on mobiles. Study of
pathogenic microbes from switches especially on fan Air conditioner (special element lead or
cadmium found in room having AC) switch in summer. The constant handling of the mobile
phones by users makes it a breeding place for transmission of microorganisms [5, 2].
Growing evidences have indicated that contaminated fomites or surfaces play a key role in
[6, 7, 8, 9]
the spread of bacterial infections . The vast majority of mobile phones are hand-held
[10]
. Combination of constant handling with the heat generated by the phones creates a prime
breeding ground for many microorganisms that are normally found on the skin.
Mobile phones are known to serve as habitats and breeding grounds for microorganisms, as
they provide a direct contact with human body and produce the heating effect while making
long calls. Mobile phones are dispensable accessories in social life but are normally not
cleaned properly. Mobile phones have also been reported to be a reservoir for
microorganisms. It has been reported that a mobile phone can harbor more microorganisms

179 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

than a mans toilet seat; the sole of a shoe or the door handle [11].
Mobile phones could be contaminated through sources such as human skin or hand, bag,
phone pouch, bags, pockets, environment and food particles, these sources are links through
which microorganisms colonized the phone, thus causing diseases that range from mild to
chronic. Although, microorganisms isolated so far by health researchers are mostly normal
flora of the source of contamination, they may serve as mobile reservoirs of infection,
allowing the transportation of the contaminating bacteria to many different clinical
environments [12].
Further, sharing of mobile phones between people may directly facilitate the spread of
potentially pathogenic bacteria to the community. The use of mobile phones by teaching, non-
teaching staff and students may serve as a potential vehicle for the spread of pathogenic
microorganisms. The constant handling of the mobile phones by different users exposes it to
an array of microorganisms and makes it a good carrier for microbes living on each square
inch of the phones. A well-practiced infection control plan that encompasses hand hygiene,
environmental decontamination, and surveillance contact isolates is effective for prevention
of such pathogenic organisms. Colonization by potentially pathogenic organisms on various
objects such as duster, marker, pen, chalk, pagers, computer, keyboards and mobile phones
has been reported and these materials are implicated in transmission of pathogen [13].
In recent time, there has been an increase in use of mobile phones by teaching, non-teaching
staff and students of educational institutions. Innovation in mobile phones has led to better
strategic life with good communication. Therefore, the use of mobile phones in the course of
working day has made mobile phones potential agents of microbial transmission. The
increase use of mobile phones is seen as responsible for rise in community infection. Hand
washing may not usually be performed often enough and many people may use personal
mobile phones in the course of a working day, the potential act of mobile phones as a source
of microbial transmission is considerable. There is no awareness in the general public of their
potential to be a reservoir of specific bacteria. The constant handling of the phone by users in
all places and occasions makes it open for arrays of microorganisms, making it a harbor and a
breeding ground for microbes especially those associated with the skin and working
environment [4].
The specific features of mobile phones having depressions at the area of key pad and other
places make it a heaven of loads of micro-organisms. From the mobile phones, different
microorganisms can spread to the other places and people. The combination of regular
handling and the heat generated by the phones creates a prime breeding ground for all sorts of

180 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

microorganisms that are normally found in our skin and environment. The human body
surface is constantly in contact with environmental microorganisms and become readily
colonized by certain microbial species. If a hand-phone microbiome connection holds for
entire bacterial communities, mobile phones can potentially be a non-invasive way to track
environmental microbial exposure over time and space, and inform how we exchange human
microbiota with our immediate surroundings. The adult human is covered with approximately
2 m of skin, with surface area supporting about 10 bacteria. The normal Microbiota is
harmless and may be beneficial in their normal location in the host but it can produce disease
if introduced into foreign locations or compromise host [14].
The distribution of mobile phones from one person to another person especially workers
working in college make possible to spread pathogenic organisms. Due to basic need of cell
phones may transfer from person to person, may be from unhygienic person to another
person. Hence, handling of mobile phones by different users exposes it to collection of
microorganisms. Because of different person handling makes a good carrier for microbes
particularly those associated with the skin resulting in the spread of different microorganism
from person to person. There are almost chances of transmission of pathogenic organisms
especially from non-teaching staff because there are number of microbes present in the
working area. Mobile phones can provide another source of information to their owners:
sample data on their personal microbiome. The personal microbiome, here defined as the
collection of microbes associated with an individuals personal effects (i.e., possessions
regularly worn or carried on ones person), likely varies uniquely from person to person.
Research has shown there can be significant interpersonal variation in human microbiota,
including for those microbes found on the skin. We hypothesize that this variation can be
detected not just in the human microorganisms, but also on the phone microorganisms [15].
The mobile phone is a personal effect so regularly carried it has been described in popular
culture as an extension of self. The reality that many people take their phones with them
everywhere they go suggests that, at any given point in time, phones and their owners are
exposed to similar environmental microbiota, which can lead to overlapping microbiota
composition. The frequency with which people directly touch their phones provides an
additional mechanism leading to shared microbiota composition. Research has shown that
traces of human microbiota are left in rooms we occupy and on surfaces we touch.If a hand-
phone microorganism connection holds for entire bacterial communities, mobile phones can
potentially be a non-invasive way to track environmental microbial exposure over time and
space, and inform how we exchange human microbiota with our immediate surroundings.

181 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

Staphylococcus aureus can cause illnesses from pimples and boils to pneumonia and
meningitis and is a close relative of methicillin resistant Staphylococcus aureus (MRSA). The
main reservoir of Staphylococcus aureus is the hand from where it is introduced into food
during handling and preparation [16].
Hand serves as a major vehicle of transmission of various microbes including the
[17]
Enterobacteriaceae group . They stressed that the microbial population of the hand is
extremely complex and variable, consisting of gram positive organisms like and
Staphylococcus aureus gram-negative organism like Pseudomonas aeruginosa, which may
survive for sufficient period of time on the hand and may thus serve either as a reservoir or
shelter of infection. Staphylococcus aureus, Enterococcus feacalis, Pseudomonas aeruginosa,
Escherichia coli, Klebsiella spp., Serratia spp., Proteus Vulgaris and Bacillus spp. were
frequently isolated from the mobile phone of health workers, marketers, food vendors,
[18, 19, 20]
lecturer, students etc . Communicable diseases like diarrhea (23%), respiratory
diseases (19%) and urinary tract infection most commonly caused by Escherichia coli,
[21, 22]
Klebsiella spp. and Staphylococcus spp. are the main disease . Cell phone of student,
teaching and non-teaching staff carries nosocomial pathogens which cause every form of skin
infections, diarrhea, Pneumonia and Meningitidess [23]. According to the WHO, antimicrobial
resistance is one of the worlds most serious public health problems.

MATERIALS AND METHOD

Study area and design

The present study was conducted entirely in the Microbial Biotechnology Lab of Post
Graduate Govt. College for Girls, Sector-42, Chandigarh. Where mobile phones were
randomly sampled from the following categories-
1. Teaching staff
2. Non-teaching staff
3. Students
The period of study was from February, 2015 and may, 2015.

Material required-Cotton swab, Mobile phones, Petri plates, Normal saline, Peptone
water, Nutrient agar, Mac Conkey agar, Sheep Blood agar, Mullor hinton agar, Imvic kit---28
kits, Gram stain kit, Antibiotic discs, Test tubes, Glass slides, Microscope, Laminar Air Flow,
Dehydrated alcohol, Oil Immersion, Test tube holder rack, Beakers, Lab cupboard, Fridge

182 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

(Microbiology lab), Measuring cylinder, Inoculating loop, Disposable Petri plates, Weighing
balance machine etc.
Isolation of bacteria from covered and uncovered mobile phones
Samples were collected from three categories: 15 teaching staff, 15 non- teaching staff, and
15 students, in which 10 teaching staff, 08 non-teaching staff and 12 students mobile phones
are covered mobile phones and 05 teaching staff, 07 non-teaching staff and 03 students
mobile phones are uncovered mobile phones from PGGCG-42, Chandigarh. The sterile swab
sticks were dipped in sterile saline and subsequently rubbed on both surface of mobile
phones. The swab was immediately inoculated into tubes containing 3ml peptone water and
incubated at 37C for a time period of 24 hrs.
Media
Preparation of peptone water/litre
Peptone 10 g
Sodium Chloride 5.0 g

Preparation of nutrient agar medium/litre

Beef /Yeast extract 1.0 g

Peptone 2.0 g
Sodium chloride 5.0 g
Agar 15 g

Preparation of Mac-Conkey Agar medium/litre

Enzymatic Digest of Gelatin 17 g
Enzymatic Digest of Casein 1.5 g
Enzymatic Digest of Animal Tissue 1.5 g
Lactose 10 g
Bile Salts Mixture 1.5 g
Sodium Chloride 5.0 g
Neutral Red 0.03 g
Crystal Violet 0.001 g
Agar 13.5 g

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Preparation of sheep blood agar medium/litre
Enzymatic Digest of Casein 0.5 g
Enzymatic Digest of Animal Tissue 0.8 g
Yeast Enriched Peptone 10 g
Corn Starch 0.1 g
Sodium Chloride 0.5 g
Agar 14 g
Sheep blood 50 ml

Preparation of Mullor Hinton Agar medium/litre

Beef Extract 0.2 g
Acid Hydrolysate of Casein 17.5 g
Starch 1.5 g
Agar 17 g

All the components given above were added in conical flasks and completely dissolved. The
pH was checked using pH stripes and adjusted using NaOH/HCl. The conical flasks was
plugged with cotton and sterilized by autoclave. The media was sterilized at 121oC at 15 psi
for 15-20 minutes.

Streak plates
The Nutrient Agar, Mac-Conkey agar and sheep Blood Agar was prepared in 250 ml flask
and was sterilized by autoclaving at 121C at 15 psi for 20 minutes. 20 ml of the media was
poured in the petri-plates before getting solidified. The media was allowed to solidify. The
inoculums of over-night peptone water were streaked on the Nutrient Agar, Mac-Conkey
Agar and sheep blood Agar medium. The petri-plates were incubated at 37C for 24 hours.
The plates were then observed for the presence of isolated colonies. After the organism
attains its full growth it was stored in refrigerator (4C) for further analysis.

Identification of bacteria
Gram staining
Gram staining is an empirical method used to classify bacterial species into two large group

184 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

gram positive and gram negative cocci and bacilli based on the chemical and physical
properties of their cell walls. The procedure is based on the ability of microorganisms to
retain purple colour of the crystal violet when decolorize with alcohol. Gram negative
bacteria are decolorized by the alcohol, losing the purple colour of crystal and remain purple.
After decolourization, safranine, a red counter stain, is used to impact a pink colour to the
decolourizer gram negative organisms. A loopful of organism was placed on a clean
microscope slide. A thin smear was prepared on a clean microscopic slide. Heat fixation was
carried out showing the slide in flame. The slide was flooded with crystal violet for 1 minute.
The slide was rinsed with tap water. A few drops of Grams Iodine was added to cover the
smear and allowed to react for 30 seconds. The slide was decolorized with 95% ethanol and
rinsed with water. The slide was flooded with safranine and allowed to react for one minute
and rinsed with tap water. The slide was blot dried and observed under oil immersion
objective.

Negative staining
Negative staining is an excellent way to determine an organisms cellular morphology. Since
the cells themselves are not stained, their morphology is not distorted in any way. The
nigrosine provides a dark background against which the shapes of the unstained cells are
clearly visible. This method provides a high degree of contrast not available in most other
staining procedures. Place a single drop of nigrosine on a clean microscope slide, adjacent to
the frosted edge. Using a flamed loop and sterile technique, remove some organism from
your tube or plate and mix it into the drop of nigrosine. Be sure there are no large clumps of
organism, but try to avoid spreading the drop. Place the end of another clean microscope slide
at an angle to the end of the slide containing the organism and spread the drop out into a film.
This is done by contacting the drop of nigrosine with the clean microscope slide and using the
capillary action of the dye/microscope slide to spread the nigrosine across the smear.. Allow
the film to air dry. Observe the slide under the microscope, using proper microscope
technique.

Biochemical analysis

Various Biochemical tests such as Indole Test, Methyl Red Test, Voges Proskauer Test,
Citrate Utilization Test, glucose, lactose, sucrose, sorbitol, mannitol, rhaminose, adonitol, and

185 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

arabinose utilization test are strike on the Imvic kit (Himedia) which was commercially
available and incubate at 37C for 18-24 hours. Next day observe the result and identify the
organism according to the biochemical reactions. Catalase Test, Oxidase Test and Coagulase
Test were also conducted to identify the isolated microorganisms.

Antimicrobial Sensitivity test

Kirby-Bauer antibiotic testing (KB testing or disk diffusion antibiotic sensitivity testing)
which is a test which uses antibiotic-impregnated disc to test whether particular bacteria are
susceptible to specific antibiotics was carried out. Fresh culture was used for Antibiotic
Sensitivity Test (AST). Colonies were transferred into 5 ml of peptone water and was
incubated at 37C for 5 hours. After that a sterile cotton swab was dipped into the sterilized
inoculum and the soaked swab was rotated firmly against the upper inside wall of the tube to
remove the excess fluid. The entire surface of the agar plate was spread properly with the
swab. Then the inoculum was allowed to dry for 10 min with the lid in place. The
predetermined battery of antimicrobial discs was dispensed aseptically onto the surface of the
inoculated agar plate. Each disc was pressed down to ensure complete contact with the agar
surface and was incubated at 37C for 24 hours. If the bacteria are susceptible to a particular
antibiotic, an area of clearing surrounds the disc where bacteria are not capable of growing
(called a Zone of Inhibition) would be observed. This along with the rate of antibiotic
diffusion is used to estimate the bacteria's sensitivity to that particular antibiotic.

RESULTS AND DISCUSSIONS

Mobile phones due to their personal nature and proximity to sensitive parts of our bodies in
usage such as face, ears, lip and hands of users could become veritable reservoirs of
pathogens that could result in infections. Results from this study showed high levels of
bacterial contamination of mobile phones used by three different categories. Specially high
levels of contamination was seen in teaching staff as compared to non-teaching staff and
students which is found significant.
Sterile swabs moistened with sterile water were rolled over the surface of both sides of the
phones. After overnight culture, all inoculated samples become turbid which indicated the
presence of bacteria in the respective samples (fig: 1). Several colonies appeared in agar plate
(Fig: 2) which differed in their size and shape (Table 5). Colonies were selected and subjected
for further gram staining and biochemical tests. The results of gram staining were identified

186 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

in different categories as gram positive cocci (Fig: 3a), gram positive (Fig: 3b) and negative
bacilli (Fig: 3c) (Table 1 & Fig: 3).

Fig: 1 Inoculated sample tubes showing turbidity

Fig: 2 isolated colonies on agars plates of different categories

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 Table1: Gram staining according to different categories showing percentage
Grams reaction Teaching staff Non-teaching staff Students Percentage
Gram positive cocci 06 06 06 40.0%
Gram negative cocci -- -- -- --
Gram positive bacilli 02 02 02 13.3%
Gram negative bacilli 07 07 07 46.7%
Total 15 15 15 100%

Fig: 3a Fig: 3b Fig: 3c

Fig: 3 Gram staining sample isolated from teaching staff

In gram staining diplococci (Fig: 4a) were seen which were gram positive cocci and were
identified by negative stain i.e. capsular stain in which capsule were seen as a clear halo
around bacteria (Fig: 4b).

Fig: 4a Fig: 4b

Fig: 4 Diplococci and capsular stain showing capsule in student sample

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 Further gram positive and negative bacteria were identified with the help of biochemical test.
Gram negative Enterobacteriaceae species were identified by HiIMViCTM Biochemical test
kit which includes four biochemical test and gram bacteria was also identified by oxidase;
catalase and coagulase test (Table 2).

Table: 2 Biochemical test for identification of an organisms

Biochemical tests 1 2 3 4 5 6 7 8 9
Indole +ve -ve -ve -ve -ve -- -- -ve --
MR +ve +ve -ve -ve +ve -- -- -ve --
VP -ve -ve +ve -ve +ve -- -- -ve --
Citrate -ve +ve +ve +ve +ve -- -- -ve --
Glucose +ve +ve +ve -ve +ve -- -- -ve --
Adonitol -ve +ve +ve -ve -ve -- -- -ve --
Arabinose +ve -ve +ve -ve -ve -- -- -ve --
Lactose +ve +ve +ve -ve -ve -- -- +ve --
Sorbitol +ve +ve +ve -ve -ve -- -- -ve --
Mannitol +ve +ve +ve +ve +ve -- -- -ve --
Rhamnose -ve +ve +ve -ve -ve -- -- -ve --
Sucrose -ve +ve +ve -ve +ve -- -- +ve --
Oxidase -- -- -- +ve -- -ve -ve -ve -ve
Catalase -- -- -- +ve +ve +ve -ve +ve -ve
Coagulase -- -- -- -ve -- +ve -ve -ve -ve
Organism E.coli Citro Entero Pseu Serra Staphyl Strep Baci Pneu
identified bacter bacter domo tia occous tococ llus mocco
nas cus cus
KEY: MR=Methyl red, VP=Voges Proskauers, - = Negative, + = Positive

189 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

 Fig: 5 Biochemical tests kits

The analysis revealed that most abundant and predominant genre of microorganisms were
Staphylococcus species 11(24.4%), E.coli 7(15.5%), Streptococcus species 6(13.3%),
Bacillus species 6(13.3%), Citrobacter species 4(8.9%), Enterobacter species 4(8.9%),
Pseudomonas species 3(6.7%), Serratia species 6 (6.7%) and Pneumococcus 1 (2.3%). Most
of them compose of the natural dermal microflora and belong to opportunistic pathogens.
Nonetheless, high amounts in the host's organism with chronic or temporary immune-
suppression can be hazardous for the cell phone owner. The mobile phones were also shown
to harbor pathogenic bacteria Staphylococcus spp. The recovery rate was between 2.3% and
24.4% (Table 3).
Table- 3 Bacteria isolated from Mobile Phones
Isolates Number recovered Percentage (%)
Staphylococcus sp. 11 24.4%
Escherichia coli 07 15.5%
Streptococcus sp. 06 13.3%
Bacillus sp. 06 13.3%
Citrobacter sp 04 08.9%
Enterobacter sp 04 08.9%
Pseudomonas sp 03 06.7%
Serratia sp. 03 06.7%
Pneumococcus sp. 01 02.3%
Total 45 100%

190 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

Pathogenic bacteria such as Staphylococcus spp, Escherichia coli, Streptococcus spp, and
Bacillus spp. were recovered in most mobile phones screened while Pneumococcus spp.
showed the least consistency. Mobile phones of teaching staff had the largest variety of
bacteria whereas non-teaching staff and students phones had the least array of bacteria.
Majority of pathogenic bacteria were isolated from mobile phones of teaching staff
(Staphylococcus species, Escherichia coli) (Table 4).

Table 4 Distribution of Bacteria Isolates from Mobile Phones according to different

Categories

Isolates Teaching staff Non-teaching staff Students

Staphylococcus spp. 05 04 02
Escherichia coli 03 02 02
Streptococcus spp. 01 02 03
Bacillus spp. 02 02 02
Citrobacter spp. 01 02 01
Enterobacter spp. 02 01 01
Pseudomonas spp. 01 02 --
Serratia spp. -- -- 03
Pneumococcus spp. -- -- 01

Gram positive cocci, Staphylococcuss, Streptococcus and Pneumococcus spp. were identified
based on morphological characteristics(Table 5), gram staining, negative staining and
biochemical tests(Fig:5). In Nutrient agar Staphylococcus spp. colonies was seen as circular,
large, shiny, smooth and pin head appearance, whereas on Blood agar haemolytic colonies
was seen and on MacConkey agar lactose fermenting colonies was observed. In gram staining
Staphylococcus spp. were seen as gram positive cocci and seen in clusters which were purple
in colour. In biochemical test Staphylococcus spp. was catalase and coagulase positive (Table
2). Staphylococcus spp. can cause food poisoning, staphylococcal scalded skin syndrome,
toxic shock syndrome. Streptococcus spp. on Nutrient agar, the colonies was seen as creamy,
circular, pin point appearance whereas on Blood agar the haemolytic colonies was seen and
on MacConkey agar non lactose fermenting colonies was observed. Streptococcus spp. was
also seen gram positive cocci and seen in chain in gram staining, which was purple in colour.
In biochemical test Streptococcus spp. was catalase, coagulase and oxidase negative (Table

191 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

2). Streptococcus spp. can cause respiratory tract infections, skin infections, genital
infections, acute rheumatic fever and acute glomerular-nephritis.
Colonies of Pneumococcus spp. on nutrient agar creamy, circular, small, mucoid and
transparent colonies whereas on Blood agar haemolytic colonies was seen and on
MacConkey agar non lactose fermenting colonies was observed. Pneumococcus spp. was
gram positive diplococci seen in gram staining (Table3a). For Pneumococcus spp. capsular
stain was done in which capsule were seen halo around the bacteria (fig3b). Pneumococcus
spp. was capsulated. In Pneumococcus spp. catalase, coagulase and oxidase was negative
(Table 2). Pneumococcus spp. can cause respiratory infections, meningitis, and bacteraemia.
The highest prevalence of Staphylococcus spp. was observed teaching staff (05) and lowest in
students (02) mobile phones. Streptococcus spp. was frequently isolated from different
categories of college and the highest rate of contamination was recorded in students (03).
Pneumococcus spp. was only observed in students (01) (Table 4).
Gram negative Bacilli, E.coli, Citrobacter, Enterobacter, Pseudomonas and Serratia species
were identified based on morphological characteristics(Table 5), gram staining and
biochemical tests(Fig:5). On Nutrient agar, E.coli colonies was seen as circular, raised, small,
smooth surface, grey-white colonies, opaque whereas on Blood agar haemolytic colonies was
seen and on MacConkey agar lactose fermenting colonies are observed (Table 5). E.coli was
gram negative bacilli and in gram staining seen as gram negative rods, which were pink in
colour and were indole, MR was utilized by bacteria and ferment only glucose, arabinose,
lactose, sorbitol, mannitol sugars(Table 2). E.coli can cause Urinary tract infections,
diarrhoea, pyro-genic infections, and septicaemia. Citrobacter spp. on Nutrient agar, smooth,
convex, non-pigmented colonies whereas on Blood agar non haemolytic colonies and on
MacConkey agar lactose fermenting colonies were observed (Table 5). In gram staining gram
negative rods were seen which was pink in colour. In biochemical tests bacteria only utilized
MR and citrate whereas bacteria only ferment glucose, adonitol, lactose, sorbitol, mannitol,
rhamnose, and sucrose (Table 2). Citrobacter spp. can cause UTI, gall bladder infections,
middle ear infections, meningitis. Enterococcus spp. was seen as gram negative rods in gram
staining, which is pink in colour. On Nutrient agar Shiny, entire margin, convex colonies
whereas on Blood agar non haemolytic colonies and on MacConkey agar lactose fermenting
colonies are observed (Table 5). In biochemical tests VP, citrate was utilized by bacteria and
ferment all sugars (Table 2). Enterobacter spp. can cause UTI, sepsis.
Serratia spp. was seen as gram negative rods in gram staining. On Nutrient agar, mucoid,
entire margins, white and red colonies appear whereas on Blood agar non haemolytic

192 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

colonies and on MacConkey agar non lactose fermenting colonies are observed (Table 5).In
biochemical tests bacteria utilized only MR, VP, citrate and only ferment glucose, mannitol
and sucrose.(Table 2). Serratia spp. can cause UTI, respiratory tract infections, meningitis,
wound infections, septicaemia, endocarditis, endotoxin shock. Pseudomonas spp. was gram
negative bacilli and seen as gram negative rods in gram staining which is pink in colour. On
Nutrient agar irregular, smooth, oval, large and pigmented colonies are seen whereas on
Blood agar haemolytic colonies and on MacConkey agar non lactose fermenting colonies are
observed (Table 5). In biochemical tests bacteria utilized only citrate and ferment only
mannitol and also catalase and oxidase positive (Table 2). Psudomonas spp. can cause UTI,
wound infections,eye infections, septicaemia infantile diarrhoea and sepsis. In gram negative
bacilli the highest prevalence of E.coli and Serratia spp. were observed. In gram staining
gram negative rods were seen. The highest prevalence of Serratia spp. was observed only in
students (03). The other gram negative bacilli bacteria were least recorded in different
categories (Table 4). Gram positive bacilli, Bacillus spp. were identified based on
morphological characteristics (Table 5), gram staining and biochemical tests (Fig: 5). Bacillus
spp. was positive rods which were purple in colour. Colonies on Nutrient agar dry, flat,
irregular, opaque, grey white colonies whereas on Blood agar haemolytic colonies and on
MacConkey agar no colonies were formed (Table 5). Bacillus spp. can cause zoonotic disease
and food poisoning. The highest prevalence of Bacillus spp. was observed in all categories
i.e. teaching, non-teaching staff and students (Table 4).

193 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

 Table 5: Cultural and Morphological characteristics of isolated organisms
Sr Organism isolated Mac-Conkey Blood Agar Nutrient Agar
no. Agar
01. Staphylococcus Lactose Haemolysis Circular, large,
spp. fermenting colonies smooth, shiny, pin
colonies head appearance

02. Escherichia coli Lactose Haemolytic Circular,raised,smal

fermenting colonies l, smooth surface,
colonies grey-white colonies,
opaque
03. Streptococcus spp. Non lactose Haemolytic creamish, circular,
fermenting colonies pin point
colonies appearance
04. Bacillus spp. No colonies found Haemolytic Dry, flat, irregular,
colonies opaque, grey white
colonies
05. Citrobacter spp. Lactose Non haemolytic smooth, convex,
fermenting colonies non pigmented
colonies colonies
06. Enterococcus spp. Lactose Non Shiny, entire
fermenting Haemolytic margin, convex
colonies colonies colonies
07. Pseudomonas spp. Non Lactose Haemolytic Irregular, smooth,
fermenting colonies oval, large and
colonies pigmented colonies.
08. Serratia spp. Non Lactose Non Mucoid, entire
fermenting Haemolytic margins, white and
colonies colonies red colonies appear
09. Pneumococcus spp. Non lactose Haemolytic creamy, circular,
fermenting colonies small, mucoid and
colonies transparent colonies

194 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

Results for the antibiotic susceptibility testing of the isolates indicated that Staphylococcus
species had the highest sensitivity of 06 against norfloxacin while E.coli had 04 against
cephotaxime whereas Streptococcus species had 05 against tetracycline and Citrobacter
species had 02 against ampicillin. On the other hand, Bacillus and pneumococcus species was
not susceptible to ampicillin, cephotaxime. Overall susceptibility was highest in nofloxacin
with 30 while the least was recorded against ampicillin with 2. In addition, the results (Table
6) show that generally, the bacterial isolates were moderately susceptible to cephotaxime 15
while higher susceptibility was obtained against norfloxacin with 30 respectively.

Fig:6 Zone of inhibition on Muller Hinton agar

195 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

 Table 6: Antibiotic susceptibility test of the bacterial isolates
Bacterial Isolate Number Amp Tet Cep Nor Kan
isolated
Staphylococcus spp. 11 -- 04 02 06 02
Escherichia coli 07 -- 02 04 05 01
Streptococcus spp. 06 -- 05 02 04 03
Bacillus spp. 06 -- 02 -- 04 02
Citrobacter spp. 04 02 01 03 02 01
Enterobacter spp. 04 -- 02 01 03 01
Pseudomonas spp. 03 -- 01 02 02 02
Serratia spp. 03 -- 01 01 02 03
Pneumococcus spp. 01 -- 01 -- 01 01
Total 45 02 19 15 30 16
KEY: Amp=Ampilcillin, Tet=Tetracyclin, Cep=Cephotaxime, Nor=Norfloxacin,Kan=Kanamycin

CONCLUSION

The overall inference of the results is that the mobile phones, though, is made to make
communication easy and accessible by almost all, and is gradually assuming the status of
pathogenic agent of disease transmission. The mobile phones examined were loaded with
large numbers of bacteria including potential disease-causing ones. The results therefore
highlight the health implications of using other peoples phones as they could be loaded with
microorganisms capable of causing various diseases and infections. Thus, cell phones could
serve as vehicles of diseases. Due to low awareness among the general people, like teaching
staff, non-teaching staff and students, personal items like mobile phone is infrequently
disinfected but extensively used everywhere even in sterile environment. Currently, there are
no restrictions for bringing or using the mobile phone within the laboratories, class rooms,
lecture halls or other places. Although, it seems impossible, in the light of all these findings,
we should be aware of limiting the mobile phone usage as it has high risk for spreading
infections. According to these results, it is obvious that, the training of teaching staff, non-

196 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017

teaching staff and students, about strict infection control procedure, hand hygiene,
environmental disinfection, and eventually, optimum disinfection methods are of great
importance. The present studies emphasize the importance of increased personal hygiene and
surface disinfection of mobile phone of teaching staff, non-teaching staff students of
PGGCG-42. Regular surface disinfection of personal items can contribute to the reduction of
transmission of pathogenic microorganism in the healthy family members.

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199 Corresponding Author- Jaspreet Kaur| Copyright | IJSS | 2017