ISSN 2320-5407 International Journal of Advanced Research (2016), Volume 4, Issue 3, 467-472

Journal homepage: http://www.journalijar.com INTERNATIONAL JOURNAL

OF ADVANCED RESEARCH

RESEARCH ARTICLE

ISOLATION, IDENTIFICATION AND CONTROL OF BACTERIA AND FUNGAL MICROORGANISMS

FROM CONTAMINATED CURRENCY NOTES.

SUBASHINI.G1, BHUVANESWARI.S2, CHITRA DEVI. K3, VIJAYALAKSHMI.R4.

1. Asst. Professor, Deparment of Microbiology, Shrimathi Indira Gandhi College, Thruchirapalli -2.
2. Asst. Professor, Deparment of Microbiology, Shrimathi Indira Gandhi College, Thruchirapalli -2.
3. Asst. Professor, Deparment of Microbiology, Shrimathi Indira Gandhi College, Thruchirapalli -2.
4. Deparment of Microbiology, Shrimathi Indira Gandhi College, Thruchirapalli -2.

Manuscript Info Abstract

Manuscript History: Money is very important to human life as it facilities the needs and currency
notes are vital for goods and services worldwide. Paper currency is used
Received: 14 January 2016
Final Accepted: 19 February 2016 repeatedly in exchange for goods and service and this are way the circulation
Published Online: March 2016 of paper currency from one individual to another potentially spreads
microorganisms. Contaminated different paper currency note samples were
Key words: collected from hospital in Tiruchirappalli,Tamilnadu. The samples were
Paper currency, contamination, analysed in microbiologically. Both gram positive and gram negative
antibiotic sensitivity test, bacteria, bacteria were found on currency notes. Predominant bacteria found in 25
fungi.
currency notes were Streptococcus pneumonia present (36%), Bacillus
subtilis (24%), Pseudomonas aeruginosa (18%), Escherichia coli (12%) and
*Corresponding Author Klebsiella pneumonia (10%). fungi were Aspergillus flavus (4%),
Aspergillus fumigatus (8%), Aspergillus niger (4%) and Candida albicans
SUBASHINI.G. (8%). DNA was separated by Agarose Gel Electrophoresis. The size of the
DNA measured using molecular marker. The bands found at 9500 bp and
8000 bp respectively. The sensitivity tests were performed to detect the
sensitivity of organisms against Standard disc placed. The maximum zone
was observed in Escherichia coli and Streptococcus pneumonia against
commercial antibiotics such as Chloramphenicol, Erythromycin .In fungi, the
maximum and minimum zone of inhibition was observed in antibiotic
clotrimazole and Amphotericin B respectively. The maximum level of
inhibition was present in 20 minutes UV treatment. Paper currency is
commonly contaminated with microbes and this may play a role in the
transmission of potentially harmful organisms. So cash should not be
handled by children and should be kept away from food and cosmetics.

Copy Right, IJAR, 2016,. All rights reserved.

Introduction:-
Microbial contaminants may be transmitted, either directly, through hand –to –hand contact, or indirectly via food or
other inanimate objects. These routes of transmission are of great importance in the health of many populations in
developing countries, where the frequency of infection is a general indication of local hygiene and environmental
sanitation levels. (Anderson, 1991; Struthers and Westran, 2003). Currency notes might act as environmental
vehicles for the transmission of potential pathogenic microorganisms (Brady and Kelly 2000). Paper currency is
used repeatedly in exchange for goods and service and this are way the circulation of paper currency from one
individual to another potentially spreads microorganisms. If these currencies recontaminated by pathogenic bacteria,
the rate of infectious diseases will continue to rise. Paper/polymer currency notes and coins may harbour various
deadly pathogenic microorganisms. Currency in the form of notes and coins represents a universal medium for the
transmission of bacteria in the environment and among humans (Xu et al.,2005). Paper currency, can be

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contaminated by droplets during coughing, sneezing, touching with previously contaminated hands or other
materials and placement on dirty surface. Pathogenic bacteria that may survive on the currency banknotes may serve
as a potential source of enteropathogens causing food poisoning because food vendors handle and serve food and at
the same time handle currency banknotes as they sell (Lamichhane et al.,2009).

Materials and methods:-

Sample collection
Contaminated 25 different paper currency note of different denominations such as 5, 10, 20.50,100 were collected
from hospital in Tiruchirappalli,Tamilnadu. They were collected in sterile polyethylene bag and transported to the
laboratory.

Isolation of microbes:-
Each currency note was aseptically transferred into individual 10 ml of sterile nutrient broth containing glass bottle
for 24 Hours. The currency was removed and the resulting nutrient broth served as a test sample. The test samples
were spread over Nutrient agar, Macconkey agar, Blood agar and PDA medium. All inoculated media were
incubated aerobically at 24 hrs (37ºC) and PDA plates were incubated at 27 ºC for 72 hrs.

Bacteriological analysis:-
Isolation of various bacterial contaminants from currency notes was performed via standard techniques
(Gilchrist,1993; Singh et al., 2002) and identified by assessing colony characteristics and Gram reaction ,Motility
test and conducting biochemical tests including for Indole production ,Citrate utilization, Urease activity ;Triple
sugar iron tests ,gas and hydrogen production tests, Catalase and Oxidase test, according to protocol described
previously (Norris and Ribbons,1972).

Identification of fungi:-
The isolated fungal species were identified by cultural character and Lacto phenol cotton blue staining. The isolated
fungal colonies aseptically transfer into clean glass slide and add one drop of Lacto Phenol cotton blue above the
mixture put a cover slip and observe the slide at high power objectives.

Extraction and separation of dna:-

DNA were isolated from contaminated bacterial and fungal cultures and separated by using Agarose Gel
Electrophoresis.

Antibiotic sensitivity test:-

The commercially available antibiotic disc such as Cephalothin, Cephoxitin, Cefuroxime, Cefixime.
Chloramphenicol, Erythromycin, Clarithromycin, Amoxycillin, Clindamycin and Vancomycin are used for bacterial
culture and Amphotericin B,Clotrimazone, Ketoconazole and Nystatin used for fungal culture.The antibiotic discs
were purchased from high media chemical Pvt. Ltd, Mumbai.The antibacterial and antifungal activities were carried
out by disc diffusion techniques. The sterile Mueller -Hinton agar plates were prepared. The bacterial and fungal
isolated organisms Escherichia coli, Klebsiella pneumoniae,Pseudomonas aeruginosa, Bacillus subtilis
Streptococcus pneumonia and Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger and Candida albicans
were spread over the Mueller -Hinton agar plates by using separate sterile cotton buds. After the microbial lawn
preparation the collected commercial antibiotic disc were placed on the organism inoculated plates with equal
distance. All bacterial plates were incubated at 370 for 24 hours. All fungal plates were incubated at 28 0C for 72
hours. The plates were observed for the zone of inhibition

Control of isolated currency microbes by UV treatment:-

The isolated microbes were treated with different interval of UV treatment. The isolates were inoculated with
nutrient broth separately. The inoculated broths were treated with UV in different intervals such as 5, 10, 15 and 20
minutes. All bacterial broths were incubated at 37 0 C for 24 hours. All fungal broths were incubated at 280C for 72
hours. After incubation all broths were read 600nm.Control also maintain (without UV treatment).

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Result and discussion:-

From the analysis of the 25 paper currency notes collected from hospital of Trichy, it was established that bacteria
and fungi were present on the notes. Totally five different bacterial colonies were isolated from the currency notes.
The isolated bacterial colonies were named as CB1CB2,CB3, CB4 and CB5.The currency note were highly
contaminated from pathogenic bacteria such as Streptococcus pneumonia present (36%),Bacillus
subtilis(24%),Pseudomonas aeruginosa(18%),Escherichia coli (12%) and Klebsiella pneumoniae(10%).Four
different fungal colonies were noted. The colonies were named as CF1,CF2, CF3 and CF4.The percentage of fungi
isolated from 25 different currency samples was denoted in( Plate–1,Table-1). Among the 25samples, 76% of
samples not found fungal growth. Aspergillus fumigatus and Candida albicans contaminated in 8% of samples
where as Aspergillus flavus and Aspergillus niger growth about 4% . Similarly results were reported (Singh and
Thakur, 2002) The other isolates found in the present study were Staphylococcus aureus(20%) and Proteus(16%)
which is approximately the same as reported in the previous reports (Gokta and Oktay, 1992) Moreover, less number
of bacteria was observed in the present study than earlier recorded

The isolated DNA from bacterial and fungal cultures were separated by Agarose Gel Electrophoresis. The results
were presented in (Plate-2, Plate-3) The size of the DNA measured using molecular marker. The bands found at
9500 bp and 8000 bp respectively. Hundred percent of the notes analyzed were found to be contaminated which is
not observed in any of the previous studies. Currency notes of lower denominations (Rs.5, Rs.10) were the most
contaminated and this is consistent with previous studies (Basavarajappa et al.,2005). This is expected, as lower
denomination notes pass through more hands than the higher denomination.

The isolated bacterial species from the contaminated currency note sample were tested for their susceptibility against
commercial antibiotics by disc diffusion method. The results were presented in (fig-1).The maximum antibacterial
activity was noted in Escherichia coli and Streptococcus pneumonia against commercial antibiotics such as
Chloramphenicol, Erythromycin, Clarithromycin, Clindamycin and Vancomycin At the same time minimum
inhibitory activity was observed against Bacillus subtilis and Pseudomonas aeruginosa. All the bacterial isolates
were resistant to Cephalothin, Cephoxitin, Cefuroxime,Cefixime and Amoxycillin. The highest antifungal activity
was noted clotrimazole against all fungal isolates. At the same time Ketaconazole highly inhibit the growth of
Aspergillus fumigates and Aspergillus niger. Moderate antifungal activity noted in Amphotericin -B and Nystatin.
The climatic and environmental conditions of the tropics favour the thriving of many pathogenic microorganisms
(Anderson,1999; Gwatkin,2000).

The isolated microbes were treated with different interval of UV treatment. The investigated results were presented
in Table-2 The maximum level of inhibition was present in 20 minutes treatments broth tubes compare than other
interval treatments
Table-1
ISOLATION OF BACTERIAL SPECIES FROM HOSPITAL CURRENCY NOTES
S.No Isolated Bacterial Strains % of occurrence
1 Escherichia coli 36
2 Klebsiella pneumoniae 24
3 Pseudomonas aeruginosa 18
4 Bacillus subtilis 12
5 Streptococcus pneumonia 10

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TABLE - II
CONTROL OF ISOLATED MICROBES BY UV TREATMENT

PLATE-1
ISOLATION OF BACTERIA

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PLATE – II
BACTERIAL DNA SEPERATION (AGROSE GEL ELECTROPHOROSIS)

PLATE – III
FUNGAL DNA SEPERATION (AGROSE GEL ELECTROPHOROSIS)

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FIG – I
ANTIBIOTIC SENTIVITY TEST AGAINST BACTERIA

Conclusion:-
Paper currency is commonly contaminated with bacteria and this may play a role in the transmission of potentially
harmful organisms. According to our results cash should not be handled by children and should be kept away from
food and cosmetics. Great care should be taken for facilitates the handling of money to avoid cross contamination.
Plastic banknotes are strongly recommended. We recommend that currency notes must be handled with caution.

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2. Anderson, R. 1999. Inhibition of Staphylococcus aureus and spheroplasts of gram negative bacteria by an
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