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The Microbiological Quality of Ice Used to Cool

Juices and Food Items in Lahore

ARTICLE FEBRUARY 2015

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2 AUTHORS:

Asif Raheem Aftab Ahmad

University of the Punjab National Academy of Young Scientists (NAY
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International Journal of Microbiology and Allied Sciences (IJOMAS)
ISSN: 2382-5537
February 2015, 1(3):19-24
IJOMAS, 2015

Original Research Article Page: 19-24

The Microbiological Quality of Ice Used to Cool Juices and Food Items in Lahore

Asif Raheem1 and Aftab Ahmad2

1
Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore, Pakistan
2
School of Biological Sciences, University of the Punjab, Lahore, Pakistan

Corresponding Author:
Aftab Ahmad
School of Biological Sciences, University of the Punjab, Lahore, Pakistan
Email: aftabac@yahoo.com

Abstract
Microbes are almost everywhere, including some of the items that we generally believe are free
from germs. Openly available ice by street vendors and fresh juice shops in Lahore city were
analyzed for their microbiological quality during the months of April to July, 2012.The total viable
counts (aerobic colony forming units (CFU)) of all 10 samples from different location of Lahore
were range from 8.8102 to 1.9105 cfu/ml. Almost all the sample collected from different places
showed high microbial load and so unsatisfactory microbiological quality. It is concluded from
results of the study that microbiology quality of ice was not good which could be due to use of
bare hands, not cleaning of utensils and quality of water used to prepare ice. The use of such ice
in food items also questions their quality so there is need of set proper standard on the quality of
ice so the consumers could be protected from bad quality food and pathogenic microorganisms.

Key words: Colony Forming Units, Ice, Pathogens, Street vendors, Sugarcane Juice.

Introduction
The World Health Organization (WHO) has
defined street vended foods as, food and
beverages prepared and/or sold by vendors in
streets and other public places for immediate
consumptions or consumption at later time
without further processing or preparation [1].
The most important vehicle of foodborne illnesses
throughout the world is contaminated food and
drinking water [2]. According to many studies, it
was illustrated that over the past decades in urban
and rural areas of developing countries,
foodborne illnesses causing high morbidity and
mortality. Lack of awareness about the food
safety, Poor and unhygienic conditions during
food preparation are the major problems in
spreading foodborne diseases in most developing
countries. Because of inexpensive, readily
available and different packages on food and
juices offered by Street vendors, it is becoming
popular among urban people as well [1]. The
standard of street venders food is very different
in developed and developing countries so street
food in developing countries is a big source of
foodborne diseases.
In most of developing countries, ice is sold by
street vendors are widely consumed by millions of
people. This is a readily available and affordable
source of ice to many sector of population
including urban poor, who cannot afford a
refrigerator at home. They take ice block from

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Asif Raheem et al. 2015
different ice factories outside city and bring those
boxes to their vendors and place at wooden table
near shops or on footpaths. Vehicles and other
transport passing nearby and dust from road are a
major source of contamination of ice. This ice is
directly used into juices from fruits and
vegetables and because of the fresh flavor,
consumer prefer unpasteurized juice. So the final
product is harbor of contamination because it is
prepared by mechanical means, contaminated ice,
punctured fruits and vegetables during harvesting
and transporting from market to vendor. Raw
materials and equipment use, traditional
processing condition, improper handling,
prevalence of unhygienic conditions all
contributes to entry of pathogenic bacteria in
juices prepared from combination of ice and fruits
[3-5]. Even if there are no pathogenic bacteria but
it often have high microbiological load. Lack of
proper information and standards in these
developing countries where street vended food is
most prevalent is a major factor contributing to
outbreaks of illnesses due to consumptions of
unpasteurized fruits juice and openly available
ice. There are several microbial studies on food in
developed and developing countries reveals
increase bacterial contamination of food with high
risk pathogens to human health. In addition, many
outbreaks were documented which are associated
with the consumption of fresh and unpasteurized
fruits juices which are prepared by vended ice [3].
In concerns with threats pose by bacterial
pathogens in juice and other food material
containing ice sold by vendors, the present work
was designed to evaluate the microbial load or
microbiological quality of vendor ice which is
used in freshly prepared juice and other food
products during April to July 2012 in different
areas of Lahore, Pakistan.
Materials and Methods
Sample collection:
Ten different ice stalls along with sugarcane
squeezed juice machine, vendors or other fruit
shops of Punjab University, Alama Iqbal Medical
College, Jinnah hospital, Barkat Market and
Moon Market, Lahore, Pakistan, were selected for
sampling. Ice samples were collected in 50ml
sterile falcon tube by wearing gloves and
International Journal of Microbiology and Allied Sciences, Feb 2015, Volume 1 Issue 3
February 2015, 1(3):19-24
transported to the laboratory in an ice box for
further processing.
Additional information were also collected and
recorded from shop owner and vendor about
delivery of ice, processing and use in food items.
Samples processing:
Sample of ice were let to fully melt at room
temperature and ice water was either used directly
or dilution was prepared with autoclaved water.
Bacteriological analysis was done through pour
plate method. 10 ml of the sample was diluted
with 90 ml of autoclaved water and mixed well (1-
10 dilution). Serial dilutions were prepared and
spread plate technique was used on enriched
media (Luria-Bertani agar (L-agar)) Sigma-
Aldarich. All plates were incubated under aerobic
conditions at 371C for 24 hrs. The mean
number of colonies counted was expressed as log
colony forming units (cfu)/ml.
Results
The samples were collected from 10 different
locations of Lahore including barkat market ice
store, hostel number 14 fruit shop (Punjab
University), barkat market Bholi juice corner,
Jinnah Hospital (sugarcane juice corner), barkat
market (ice slush maker), hostel number 15 stop
sugarcane juice vendor (canal road, Punjab
University), Allama Iqbal Medical college fruit
shop, hostel number 4 (Punjab University)
canteen fruit shop, moon market ice store and
moon market fruit shop. All the samples showed
microbial load ranges from 8.8102 to 1.9105
cfu/ml. The maximum number of bacterial load in
ice was recorded in barket market Bholi juice and
Jinnah hospital sugar cane shop (6.52103)
followed by moon market ice store (3.62103) and
minimum number of bacterial load was at barket
market ice store (8.8102) and shops in University
of the Punjab (Table. 1).
The total numbers of colonies obtained with 1:10
dilution of ice water are shown in Figure 1 with
lowest in IW-1 and highest in IW-3 and 4.
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Asif Raheem et al. 2015 February 2015, 1(3):19-24

Sample ID Location Dilution factor No. of Colonies CFU/ml

IW-1 Barkat market Ice Store 0 Numerous
10-1 88 8.8102
10-2 80 8103
10-3 70 7104
IW-2 Hostel no.14 Fruit Shop 0 310
10-1 227 2.27103
10-2 90 9103
10-3 70 7103
IW-3 Barkat Market Bholi Juice 0 Numerous
Corner
10-1 652 6.52103
10-2 225 2.25104
10-3 150 1.5105
IW-4 Jinnah Hospital (Sugarcane juice 0 Numerous
corner)
10-1 652 6.52103
10-2 270 2.7104
10-3 190 1.9105
IW-5 Barkart market (Ice Slush 0 Numerous
Maker)
10-1 320 3.2103
10-2 225 2.25104
10-3 102 1.02105
IW-6 Hostel no.15 stop Sugarcane 0 Numerous
Juice Vendor
10-1 156 1.56103
10-2 124 1.24104
10-3 75 7.5104
IW-7 Allama Iqbal Medical College 0 Numerous
Fruit Shop
10-1 228 2.28103
10-2 85 8.5103
10-3 60 6104
IW-8 Hostel no.4 Canteen Fruit Shop 0 Numerous
10-1 226 2.26103
10-2 79 7.9103

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Asif Raheem et al. 2015 February 2015, 1(3):19-24

10-3 53 5.3104
IW-9 Moon Market Ice Store 0 520
10-1 362 3.62103
10-2 162 1.62104
10-3 85 8.5104
IW-10 Moon Market Fruit shop 0 Numerous
10-1 265 2.65104
10-2 182 1.82104
10-3 122 1.22105

Table 1: Colony forming units (CFU) of all the samples collected from different places of
Lahore.

sp., Citrobacter sp. and vibrio cholera were

Figure 1: The total number of colonies from
ice sample with 1:10 dilution
Discussion
Contaminated water is a big source of
contamination of food worldwide and ice which is
generally considered germ free can also be a
source of contamination. In the present study, we
checked the general hygiene of ice by aerobic
CFU count of total aerobic microbial load in ice,
which was in use to cool juices, drinking water
and other food items in Lahore area. Lahore is 2nd
largest city of Pakistan by population. According
to studies, deadly pathogens like, Listeria
monocytogenes, E. coli, S. aureus and other
intestinal pathogens involve in variety of diseases
can be present in ice. In addition, Bacillus sp.,
Streptococcus sp., Staphylococcus
epidermidis, Enterobacter aerogenes, Klebsiella
isolated and identified from contaminated ice [3].
In an outbreak study, Florida, USA, about 60
visitors were affected due to consumption of fresh
non-pasteurized juice which was contaminated
with salmonella [6] and one source of such
contaminations in developing countries could be
ice.
According to a study conducted in Australia,
about 427 confirmed cases of salmonellosis were
reported in 1999 after drinking unpasteurized
orange juice [3]. Another study illustrated that in
India, an epidemic was reported in Pune city,
which was because of sugarcane juice containing
ice contaminated with Vibrio cholerae [7].
Numerous studies showed that microbial quality
of manufactured ice is also compromised and
could be source of contamination. Lateef et al.,
(2006) evaluated microbial safety of
manufactured ice which was used to cool food and
drinks in Nigeria and it was observed that
microbial load of these manufactured ice samples
were in the range of 1.88-3.20104cfu/ml, which
was more than that of recommended microbial
load for ice [8].
In our study, it was observed that quite high
microbial load was present and especially at
places which serve fresh juices to customers.
Sometime people get food poisoning after
drinking juice and beside many other factors
(juice squeezing machine, operator etc.) ice could
be a factor as it may contain a high dose of

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Asif Raheem et al. 2015
pathogenic bacteria and that can cause food
poisoning and other illnesses.
As there are microbiological standard for drinking
water, there should be for ice as ice is used in food
items, juices and cold drinks almost everywhere
in the world. The ice should be of microbiological
quality and it should not have high load of
microbes. The ice of satisfactory microbiology
quality should have aerobic plate count or CFU
that should not exceed 103 CFU/ml [9] but in our
case just 2 samples (IW-2 and IW-9) had
microbial count less than this, while all the other
had high quantity of microbes and so was not of
satisfactory level.
Although we could only check the aerobic CFU,
other factors are also necessary to evaluate like
presence of total coliforms and E. coli,
Enterococci, Salmonella and other related
organisms, which is true indication of quality of
ice and also have direct link with possible human
diseases. This was a small study, with limited
number of samples, a large scale and countrywide
study is needed to have better idea about quality
of ice in Pakistan. In addition, we observed that at
all the places, the vendor added ice directly by
hands so there was strong possibility of presence
of above microbes along with ice as un-cleaned
hand are great source of contamination [10].
Moreover, all the utensils which were in direct
touch with ice were not very clean and vendors do
not clean them quite often. In addition, ice was
also not properly covered with plastic or other
sheets to avoid direct contact of surrounding air.
According to microbiological standards, the ice
used in food items should be of same
microbiological quality as potable water and
E.coli and Enterococci should not be present in
100 ml [9].
The study shows that ice sold by vendors and used
in fresh fruit juices was not of good
microbiological quality as it contained high
aerobic CFU/ml, so it also put question on the
quality of food items in which it is used. The ice
utensils in which ice is stored should be cleaned
frequently and also should not be added into food
items with un-cleaned bare hands. In addition,
regulations should be set on ice quality and those
standards should be followed all over the country
to have good quality food for all the people. There
is also need of public awareness that ice which is
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February 2015, 1(3):19-24
considered as germ free is not microbe free and it
can contain potentially pathogenic bacteria which
can put their health at risk.
References
1. World health Organization (WHO). 1996.
Essential safety requirement for streets
vended foods (revised edition), food safety
unit, division of food and nutrition, WHO,
Geneva.
2. World health Organization (WHO). 1999.
Food safety: an essential public health issue
for the new millennium food safety program,
department of protection of human
environment, cluster of sustainable
development and healthy environment, WHO,
Geneva.
3. Victorian Government Department of Human
Services, Food Safety Unit Melbourne,
Victoria. 2005. Microbiological survey of
freshly squeezed juices from retail businesses
across Victoria. Available at:
http://www.health.vic.gov.au/foodsafety
(accessed September 12, 2014)
4. Oliveira ACG, Seixas ASS, Sousa CP, Souza
CWO. 2006. Microbiological evaluation of
sugarcane juice sold at street stands and juice
handling conditions in So Carlos, So Paulo,
Brazil. Cad. SadePblica, Rio de Janeiro.
22(5):1111-1114.
5. Nicolas B, Razack BA, Yollande I, Aly S,
Tidiane OCA, et al. 2007. Street-Vended
Foods Improvement: Contamination
Mechanisms and Application of Food Safety
Objective Strategy: Critical Review. Pakistan
Journal of Nutrition. 6(1):1-10.
6. Schmidt RH, Sims CA, Parish ME, Pao S,
Ismail MA. 1997. A model HACCP plan for
Small-Scale, Fresh-Squeezed (Not
Pasteurized) Citrus Juice Operations.
7. Mosupye FM, von Holy A. 1999.
Microbiological Quality and Safety of Ready-
To-Eat Street- Vended Foods in
23
Asif Raheem et al. 2015 February 2015, 1(3):19-24

Johannesburg, South Africa. Journal for Food 10. Burton M, Cobb E, Donachie P, Judah G,
Protection. 1278-1284. Curtis, et al. 2011. The effect of handwashing
8. Lateef A, Oloke JK, Kana EB, Pacheco E. with water or soap on bacterial contamination
2006. The Microbiological Quality of Ice of hands. International Journal of
Used to Cool Drinks and Foods in Ogbomoso Environmental Research and Public Health.
Metropolis, Southwest, Nigeria. Internet 8(1):97104.
Journal of Food Safety. 8:39-43.
9. Nichols G, Gillespie I, and de Louvois J. 2000.
The microbiological quality of ice used to
cool drinks and ready-to-eat food from retail
and catering premises in the United Kingdom.
Journal of Food Protection. 63(1):7882.

For Citation:
Raheem A and Ahmad A. 2015. The Microbiological Quality of Ice Used to Cool Juices and Food
Items in Lahore. International Journal of Microbiology and Allied Sciences. 1(3):19-24.

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