Int.J.Curr.Microbiol.App.

Sci (2015) 4(1): 340-347

ISSN: 2319-7706 Volume 4 Number 1 (2015) pp. 340-347

http://www.ijcmas.com

Original Research Article

Microbial profiling of street foods of different locations at
Dehradun city, India

Neha Chauhan1*, Veena Uniyal1 and Devendra Singh Rawat2

1
Department of Microbiology, Baba Farid Institute of Technology, Suddhowala, Dehradun, India
2
Department of Life Sciences Baba Farid Institute of Technology, Suddhowala, Dehradun, India
*Corresponding author

ABSTRACT

Keywords Improper personal hygiene can facilitate the transmission of the pathogenic bacteria
found in environment and on people s hands via food to humans. The present study
Personal
was undertaken to investigate the microbiological quality of different food in
hygiene,
Dehradun, India. For the microbial screening of various food samples, Pour plate
Pathogenic
technique and biochemical characterisation had been performed. Results showed
Bacteria,
the presence of considerable number of contaminating microorganisms which lead
Food
to several food borne infections. Food hawkers in India are generally unaware of
Regulations,
food regulations and have no training in food-related matters. They also lack
Supportive
supportive services such as water supply of adequate quality and disposal systems,
services
which hamper their ability to provide safe food.

Introduction

Street foods are defined as ready to eat foods who are illiterate and do not practice
and beverages sold by vendors and hawkers hygiene. The chances of contamination of
especially in the streets and other similar these foods increase due to the poor
places (FAO, 2000). Existence of Street environmental conditions in which the
foods is because of their easy availability, preparation is done and sold (Sheth, 2005).
variety, prevailing socio economic Street foods are the cause of several types of
conditions and also the influx of food - borne disease. The water used for
unorganized labour. Though the concept of drinking and cleaning purposes is often
street foods was unwelcome initially, Their contaminated due to unhygienic storage and
existence depends on certain factors such as handling. Moreover use of artificial colours,
increasing labour force and consumer like metanil yellow, has led to serious health
demand due to rapid industrialization hazards. Proper garbage removal facilities
(Chakravarthy, 1995). are also not available, thus leading to poor
environmental condition (Chakravarthy,
Safety of street foods is always a matter of 2003).
concern as in most cases they are prepared
under unsanitary conditions by the vendors Chutneys are prepared from available

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seasonal fruits and vegetables, herbs, which recognized problem involving a wide
are ground to a paste or to a pulpy mash; spectrum of illnesses caused by bacterial,
requisite consistency is obtained by addition viral, parasitic or chemical contamination of
of water, vinegar and lime or tamarind juice. food. Although viruses account for half of
The prevalence and growth of pathogens on all the food borne illnesses, most
the raw foods especially vegetables, salads, hospitalizations and deaths related to food
fruits and sprouts, are used as ingredients of borne infections are due to bacterial agents.
chutneys (Viswanathan and Kaur, 2001). Diarrheal diseases are the commonest
The microbiological qualities of chutney symptoms of food poisoning and in some
sold on the street in metropolitan cities are cases, can lead to death. The diseases are
very poor (Kakar and Udipi, 2000). Almost caused by either toxin from the disease-
all samples on street vended food possess causing microbe, or by the human body s
pathogenic organisms like Salmonella reactions to the microbe itself (Teplitski et
species, Shigella species, Campylobacter al., 2009).
species; E. coli can contaminate the food
through contact with samples. Food borne The work examined the microbiological
illness caused by eating microbial quality of different food types such as Aloo
contaminated food is an important public tikki, Momo, Chowmein and Chutney, sold
health problem. at three different locations of Dehradun.
These locations were chosen because of the
Food is contaminated by various pathogenic large number of commuters that patronize
microorganisms which cause food infection the spot on daily basis.
or food intoxication. Food poisoning can be
the result of either chemical or the ingestion Materials and Methods
of toxicant. Bacterial food intoxication
therefore refers to food borne illness caused Sample Collection
by the presence of a bacterial toxic formed
in the food (Mensah et al., 1999). This may In order to determine the availability of
be an attempt to make aware common street foods, three different locations were
people regarding microbial contamination of randomly selected from city and a survey
street vended food and health hygienic. was conducted. Various food samples (Aloo
tikki, Momo, Chowmein and Chutney) were
Despite the economic and nutritional collected from 3 different locations in
benefits of street foods, the consumption of Dehradun city.
these roadside foods has been suggested to
potentially increase the risk of food borne Microbiological analysis
diseases as street foods are readily
contaminated from different sources For the microbiological analysis of food
(Tambekar et al., 2008). In fact, street foods sample, each sample was thoroughly mixed.
have often been associated with traveller s Serial dilutions of samples were prepared for
diarrhea and other food borne diseases. further analysis. After serial dilution pour
Studies have revealed the frequent plate technique was applied. After
contamination of street food in many solidifying, Petri plates were incubated at
developing world including Nigeria. 370C for 24 hours in inverted position. After
incubation the plates with maximum number
Food borne diseases are an increasingly of colonies was selected and the number of

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colonies were counted on the selected plates. incubation, iodine solution was poured on
The isolated colonies of organism were the agar and examined for hydrolysis of
transferred to nutrient agar slant for starch by the production of clear zone
maintenance and further identification. around the microbial growth.

Identification and Biochemical Gelatin liquefaction: Gelatin is a protein

characterization of Isolates produced by hydrolysis of collagen.
Hydrolysis of gelatin is brought about by
The isolated colonies were morphologically microorganism capable of producing a
characterized e.g., colonial growth and proetolytic exoenzyme known as gelatinase.
pigmentation. In biochemical The degradation of gelatin occurs in the
characterization, Catalase test, Amylase, medium by an exoenzyme, it can be detected
Gelatine Liquefaction, IMViC test, Oxidase, by observing liquefaction (i.e. flooding the
Urease, Carbohydrate fermentation (lactose, gelatin agar medium with mercuric chloride
sucrose, dextrose) were performed. solution and observe the plates for clearing
around the line of growth) because gelatin is
Biochemical tests and Staining were done also precipitated by chemicals that coagulate
in accordance to standard procedure proteins while the end products of
(Aneja, 2003) degradation (i.e. amino acids) are not
precipitated by same chemicals.
Gram staining: The gram stain is a
differential stain which is used to Urease test: Urease test is performed by
differentiate bacteria into two groups Gram growing the test organisms on urea broth or
positive bacteria and Gram negative agar medium containing the pH indicator
bacteria. The technique is based on the fact phenol red (pH 6.8). During incubation,
that Gram positive cell wall has a stronger microorganisms possessing urease will
attraction for crystal violet when iodine is produce ammonia that raises the pH of the
applied and therefore retains the crystal medium/broth. As the pH becomes higher,
violet and therefore will remain purple after the phenol red changes from a yellow colour
decolorizing while Gram negative bacteria (pH 6.8) to a red or deep pink colour.
will be colourless after decolorizing with Failure of the development of a deep pink
alcohol, counterstaining with Safranin will colour due to on ammonia production is
make them to appear pink. evidence of a lack of urease production by
the microorganisms.
Catalase test: The glass slide was held at an
angle and few drops of 3% hydrogen Oxidase test: The oxidase test is a test used
peroxide were allowed to flow slowly over in microbiology to determine if
the culture. The emergence of bubbles from a bacterium produces certain cytochrome c
the organism was noted. The presence of oxidases. It uses disks impregnated with
bubble displayed a positive test indicating a reagent such as N, N, N , N -tetramethyl-p-
the presence of enzyme catalase. If no gas is phenylenediamine (TMPD) or N, N-
produced, this is a negative reaction. dimethyl-p-phenylenediamine (DMPD),
which is also a redox indicator. The reagent
Amylase test: In this test, isolate was point is a dark-blue to maroon color when
inoculated on starch agar plates and oxidized, and colorless when reduced.
incubated at 370C for two days. After Oxidase-positive bacteria possess

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cytochrome oxidase or indophenol oxidase Methyl red test: All enteric

(an iron-containing hemoprotein). These microorganisms ferment glucose and
both catalyze the transport of electrons from produce organic acids. The methyl red
donor compounds (NADH) to electron indicator which is used in this test, in the pH
acceptors (usually oxygen). The test reagent, range of 4 will turn red, which is the
TMPD dihydrochloride acts as an artificial indicative of a positive test. At a pH of 6,
electron donor for the enzyme oxidase. The still indicating the presence of acid but with
oxidized reagent forms the colored a lower hydrogen ion concentration, the
compound indophenol blue. The cytochrome indicator turns yellow and is a negative test.
system is usually only present in aerobic
organisms that are capable of using oxygen Voges Proskauer test: Voges Proskauer
as the final hydrogen receptor. The end- test determines the capability of some
product of this metabolism is either water or microorganisms to produce non-acidic or
hydrogen peroxide (broken down neutral end products, such as acetyl methyl
by catalase). Live bacteria cultivated carbinol, from the organic acids that results
on trypticase soy agar plates may be from glucose metabolism. The reagent used
prepared using sterile technique with a in this test, Baritt reagent consists of
single-line streak inoculation. The mixture of alcoholic alpha- napthol and
inoculated plates are incubated at 37°C for potassium hydroxide solutions. Detection of
24 48 hours to establish colonies. Fresh acetyl methyl carbinol requires this end
bacterial preparations should be used. After product to be oxidized to a diacetyl
colonies have grown on the medium, 2-3 compound. This reaction will occur in the
drops of the reagent DMPD are added to the presence of alpha-napthol catalyst and a
surface of each organism to be tested. A guanidine group that is present in the
positive test (OX+) will result in a color peptone of MR-VP medium. As a result, a
change violet to purple, within 10 30 pink complex is formed, imparting a rose
seconds. A negative test (OX-) will result in colour to the medium.
a light-pink or absence of coloration.
Citrate utilization test: In the absence of
IMViC fermentable glucose or lactose, some micro-
organisms are capable of using citrate as a
Indole test: Tryptophan is an essential carbon sources for energy. This ability
amino acid that can undergo oxidation by depends on the presences of citrate
the way of enzymatic activities of bacteria permease that facilitates transport of citrate
and converted into metabolic products in the cell. During this reaction the medium
(indole, pyruvic acid and ammonia) is becomes alkaline, the carbon dioxide that is
mediated by the enzyme tryptophanase. The generated combines with sodium and water
presence of indole is detected by adding to form sodium carbonate, an alkaline
Kovac s reagent which produces cherry red product. The presence of carbonate changes
colour. The colour is produced by the the Bromothymol blue indicator
reagent which is composed of p- incorporated into the medium from green to
dimethylaminobenzaldehyde yielding the Prussian blue. After incubation citrate
cherry red colour. Absence of red coloration positive culture are identified by the
demonstrates that the substrate tryptophan presence of growth on the surface of slants,
was not hydrolysed and indicates an indole which is accompanied by blue coloration.
negative. Citrate negative will show no growth and
the medium will remain green.
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Carbohydrate fermentation: Fermentation can facilitate the transmission of these

of carbohydrates (glucose, sucrose, lactose pathogenic bacteria found in environment
etc.) are carried out by microorganism, and on people s hands via food to humans
under anaerobic condition in which a (Tambekar et al., 2008). The present study
Durham tube is placed in inverted position was undertaken to investigate the
to trap the gas bubble formed due to microbiological quality of different food
production of gas. The fermentation broth types in Dehradun, India. Food hawkers in
contains ingredients of nutrient broth, a India are generally unaware of food
specific carbohydrate and a pH indicator regulations and have no training in food-
(phenol red), which is red at neutral pH (7) related matters. They also lack supportive
and turns yellow at or below a pH of 6.8 due services such as water supply of adequate
the production of an organic acid. quality and rubbish disposal systems, which
hamper their ability to provide safe food
Results and Discussion Titarmare et al. (2009)

Food borne Diseases are caused by wide The results obtained from various food
spectrum of illness caused by pathogenic samples which were screened for the
bacterial, viral, protozoan and chemical presence of food borne pathogens have been
contamination of food which has been discussed below. High levels of bacterial
recognized as major problem. Major cause contamination at varying degrees were
for the rapid increase in the occurrence of detected in the food types tested. Samples of
Food borne infections is the Street Foods Aloo tikki, Momo, Chowmein and chutney
which is easily affordable by poor people. had considerable levels of contamination
Food poisoning can be the result of either which can lead to various types of serious
chemical or the ingestion of toxicant. This diseases.
may be an attempt to make aware common
people regarding microbial contamination of Total of 10 bacterial isolates were obtained
street vended food and health hygienic from the different street food types out of
conditions. which 5 isolates were found to be Gram
positive and 5 Gram negative. Some of cells
Isolation and Enumeration of Isolates were arranged singly, paired or in chains or
from Food samples clusters. Some of them are coccus in shape
while others are rod in shape. They showed
The various street food samples (Aloo Tikki, different morphologic patterns on agar plate
Momo, Chowmein, and Chutney) were as circular, opaque, round, entire, pin head
screened for the presence of pathogens colonies. They also showed varied
responsible for causing Food borne coloration as white, cream, pink, gelatinous,
infections by Pour plate technique at mucoid. These unknown bacterial isolates
different dilutions. The no. of colonies were biochemically characterized by
obtained at different dilutions is compiled in different tests. E. coli and other coliform
Table 1 and graphical representation bacteria could be due to inadequate hand
showing the number of colonies (CFU/ml) washing by food workers and the absence of
in different samples at different locations is good manufacturing practices. The presence
presented in Figure 1. of S. aureus caused severe contamination
through handling (Tambekar et al., 2007).
Conclusively, improper personal hygiene

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When bacterial isolates were subjected to aureus, Bacillus cereus, Salmonella,

catalase test to detect the development of Shigella. All these bacterial pathogens are
bubbles after the addition of hydrogen responsible for the food borne and diarrheal
peroxide, isolates S1, S2, S3, S7, S8 showed diseases.
positive result. S6, S8, S10 bacterial isolates
showed positive amylase test by producing a Most food pathogens are of soil or intestinal
clear zone around the colonies i.e starch origin and are transmitted through poor food
hydrolysis reaction. Fermentative preparation, personal hygiene or public
degradation of carbohydrates such as sanitation practices. Therefore to ensure the
sucrose, lactose and dextrose was done by safety of the foods, producer and hawkers
all the isolates by producing acid and gas. must maintain a clean environment,
S1 and S3 isolates showed positive Indole minimize contact with the food samples
test while others gave negative result. When after production and also maintain a high
subjected to MR test isolates S1, S2, S3, S7, level of personal hygiene. Also, utensils
S9 showed positive MR result and S6, S7 should be properly clean at all stages of
produced positive VP test. S2, S4 isolates production.
were found to be Citrate utilization positive.
Cultures S6, S7 showed positive result for The local Government and the ministry
Gelatin liquefaction and S4, S5, S10 isolates should consider establishment of adequate
were urease positive. facilities and utility services as well as
provision of necessary information,
In the present study, the bacteriological education and training programmes for
quality of different food samples such as vendors and consumers. Our findings have
Aloo tikki, Momo, Chowmein, Chutney shown that there is need for improvement of
were found to be contaminated with good hygiene practices (GHP) so that street
different bacterial pathogens like E. coli, S. food contamination can be reduced

Table.1 Microbial count in different samples at various locations

Samples Dilution factor Number of colonies (CFU/ml)

Location 1 Location 2 Location 3
-5
Allu Tikki 10 10 30 12
10-6 8 25 8
10-7 7 15 7
Momo 10-5 Conjugated 35 32
Colonies
10-6 15 30 27
10-7 12 22 20
Chowmein 10-5 10 10 38
10-6 7 7 35
10-7 5 5 25
Chutney 10-5 35 33 62
10-6 28 30 58
10-7 20 28 35

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Fig.1 Graphical representation of microbial counting different food samples (Allu Tikki,
Momo, chowmein, Chutney) at different locations in Dehradun

References city. J. Food. Sci. Technol., 87(5):

509 51.
Aneja, K.R. 2003. Staining and Mensah, P., Owusu-Darko, K., Yeboah-
biochemical techniques. In: Manu, D., Ablordey, A., Nkrumah,
Experiments in microbiology, plant F.K., Kamiya, H. 1999. The role of
pathology and biotechnology, 4th street food vendors in the
edn. New Age International (p). transmission of enteric pathogens.
Ltd., Publishers, New Delhi. Ghana Med. J., 33: 19 29.
Chakravarthy, I. 1995. Urban street food Sheth, M., 2005. Food safety concerns:
in Calcutta. Proc. Nutr. Soc. India, Strategies to Combat. Pp. 1 10.
42: 53 59. Tambekar, D.H., Gulhane, S.R.,
Chakravarthy, I. 2003. Street foods: Jaisingkar, R.S., Wangikar, M.S.,
Safety. Risk and Nutrition Potential, Banginwar, Y.S., Mogarekar, M.R.
Nutrition Goals for Asia-Vision 2008. Household water
2020. Nutrition Foundation of India. management: A systematic study of
Pp. 660 662. bacteriological contamination
FAO, 2000. Street foods in Asia. Food between source and point-of-use.
Safety and Nutritional Aspects Am.-Eur. J. Agricult. Environ. Sci.,
Workshop. Pp. 1 4. 3(2): 241 246.
Kakar, D.A., Udipi, S.A. 2000. Tambekar, D.H., Shirsat, S.D., Suradkar,
Microbiological qualities of different S.B., Rajankar, P.N., Banginwar,
varieties of chutneys sold in Mumbai Y.S. 2007. Prevention of

346
 Int.J.Curr.Microbiol.App.Sci (2015) 4(1): 340-347

transmission of infectious disease:

Studies on hand hygiene in health-
care among students. Continental J.
Biomed. Sci., 1: 6-10.
Teplitski, M., Wrigh, A.C., Lorca, G.,
2009. Biological approaches for
controlling shellfish-associated
pathogens. Curr. Opin. Biotechnol.,
20: 185 190.
Titarmare, A., Dabholkar, P., Godbole, S.
2009. Bacteriological analysis of
street vended fresh fruit and
vegetable juices in Nagpur city,
India. Int. J. Food Saf., 11: 1 3.
Viswanathan, P., Kaur, R. 2001.
Prevalence and growth of pathogens
on salad vegetables, fruits and
sprouts. Int. J. Hyg. Environ.
Health., 20: 205 213.

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