Euphrates Journal of Agriculture Science-6 (4): 30-37 , (2014)
Jar allah et al
Isolation and Identification of some pathogenic Bacterial Species
Contaminated from Meats in Butchers Shops and Kebab
Restaurants in AL-Kut city
Eman M. Jarallah
Seaab Imad Sahib & Khalid Yassen
Collage of science \ Babylon University Technical Institute / Kut
Abstract:
This study was conductedfor detectionof pathogenicbacterial species that
contaminate meat in the butchers shops and kebab restaurants inAl-Kut city. Ten
samples from 10 butcher shops and 10 restaurantswere collected.These samples were
suspended in sterile normal saline in order to using for isolation and identification of
pathogenic bacteria. The cultural properties and biochemical tests results revealed
the bacterial isolates return to two+ bacterial species: Escherichia coli (40%) and
Staphylococcus aureus (29%) in butchers shops, in front of E.coli (19%) ,S.aureus
(28%) and Klebsiella sp. (9%) in restaurants . The antibiotics susceptibility pattern
results showed all of these isolates were resist to most traditional antibiotics but in
different ratios.
‫عسل وتشخيص تعض اصناف الثكتريا الورضية هناللحوم في هحالت القصاتة وهطاعن الكثاب في‬
‫هذينة الكوت‬
‫خالذ ياسيي زغيرالساهلي‬
‫صعاب عواد صاحة‬
‫ايواى هحوذ جار هللا‬
: ‫الخالصة‬
‫اجزيث ٌذي الذراسة للكشف عه اصىاف الثكحزيا المزضية الملُثً للحم في محالت القصاتة َمطاعم الكثاب في‬
‫ َضعث ٌذي العيىات في المحلُل الملحي‬.‫ مطاعم‬03َ ‫ محالت قصاتة‬03 ‫ عيىات مه‬03 ‫ جمعث‬.‫مذيىة الكُت‬
‫ َقذ اظٍزت الىحائج الحي جم الحصُل‬.‫الفسلجي المعقم لغزض اسحخذامٍا في العزل َجشخيص الثكحزيا المزضية‬
:‫عليٍا مه خصائص الُسط الزرعي َاالخحثارات الكيمُحيُية تان الثكحيزيا المعزَلة جعُد الى صىفيه ٌَما‬
‫) في محالت القصاتة اما في مطاعم‬%92( ‫) َالمكُرات العىقُدية الذٌثية تىسثة‬%03( ‫االشزيشياالقُلُوي تىسثة‬
‫) َالكلثسيلة‬%92( ‫) َالمكُرات العىقُدية الذٌثية تىسثة‬%02( ‫الكثاب فقذ ظٍزت االشزيشيا القُلُوية تىسثة‬
‫ اما وحائج فحص الحساسية للمضادات الحيُية فقذ اظٍزت ان جميع الثكحزيا المعزَلٍٍي مقاَمة‬.)% 2( ‫تىسثة‬
.‫الغلة المضادات الحيُية َلكه تىسة مخحلفة‬
important are water availability, pH,
and temperature (ICMSF, 1996).
Food-borne diseases are an
important cause of morbidity and
mortality worldwide. It is estimated
that
foodborne
diseases
cause
approximately 76 million illnesses,
325,000 hospitalizations, and 5,000
deaths in the United States each
year(Mead
et
al.,1999).People's
Introduction:
Food is a chemically complex matrix,
and predicting whether, or how fast,
microorganisms will grow in any given
food is difficult. Most foods contain
sufficient nutrients to support microbial
growth. Several factors encourage,
prevent, or limit the growth of
microorganisms in foods; the most
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Euphrates Journal of Agriculture Science-6 (4): 30-37 , (2014)
increased traveling and free movement
of foodstuffs has increased the risk of
contracting
food
poisonings
(Shahramet al., 2012).
Contaminated
raw
or
undercooked red meats are particularly
important in transmitting these food
borne
pathogens(Mengand
Doyle
,1998).Escherichia coliare considered
the
most
commensally
living
microorganism in the alimentary tract
of nearly all domestic and wild animals
as well as human.EnteropathogenicE.
coli organisms usually lead to severe
diarrhea in infants and it may also be
the causal organisms in appendicular
abscess, peritonitis and cholecystitis(
Mackieand McCartney,1989).
The Enterobacteriaceae group of
bacteria is the most challenging
bacterial contaminant to raw and
processed meat products worldwide.
E. coli, Klebsiellaspecies are the most
predominant speciesin all food
poisoning cases associated with some
meat products, Due to the rising
incidence of food borne infections,
there is an urgent need for control
and/or prophylaxis for food poisoning
outbreaks associated with meat
products,it
depends
greatly
on
investigating the causative agents in
food (meat products), eliminating them
to ensure food safety and to protect
public
health
from
microbial
contamination
of
food(AlMutairi,2011). Staphylococcal food
poisoning is overwhelmingly the most
prevalent of food borne infections in
the United States (Ash,1997).
This study carried out to
investigate the diseases that caused by
food contamination by the studied
pathogens and determination the most
bacteria that contaminate the fast food.
Materials and Methods:
Collection of samples:
Ten fast food restaurant and ten
meat
butcher
shopsin
Kut
citywerechosen randomly to be used
in the present study. A total of 10
samples of fresh meat and 10 Kebab
samples were obtainedaseptically and
transferred to the laboratory in sterile
plastic bagskept in Ice-Box, according
to (Cheesbrough,1984).
In the laboratory, one gram of
each sample was weighted out and
homogenized into 9 ml of sterile
distilled water. From the ten –fold
dilutions of the homogenates:0.1ml 0f
10-2 ,10-3and 10-4dilutions were plated
in culture on theNutrient broth media,
Nutrient
Agar,
Blood
Agar
andMacconky's Agar by pour method
,The plates were then incubated at 37˚C
for 24-48 hours .After the end of
incubation period , colonies were
counted using colony counter .The
count were expressed as colony
forming units(CFU). Pure isolates of
bacterial species were then stored at
4◦C for using in identification
examination.
Identification of bacterial species:
The
growing
colonies
transferred to new specialized media
for each bacteria to obtain for a pure
culture. Again the isolated bacteria
were cultured at a 37˚C for 24h
andstaining procedure were applied by
using Gram stain. The biochemical
tests were conducted by API 20 Kit to
identify the isolated bacterial species.
Antibiotics susceptibilitytest:
Kirby-Bauer disk diffusion method was
used to determine the antibioticresistant characteristics of the isolated
organisms (Bauer et al.,1996).Mueller00
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Jar allah et al
Euphrates Journal of Agriculture Science-6 (4): 30-37 , (2014)
samples
from
butcher
shops
werecontaminated with two strains of
bacteria (Table.1). Figures 1 and 2
represent the frequency of the
pathogenic bacteria in studied Kebab
and meat samples, respectively.As
shown in these figures the positive
species S.aureus was the more frequent
pathogen in Kebab samples followed
by E.coli and finally Klebsiella sp.
While in meat samples E.coli came at
the first then S.aureus .
The antibiotics susceptibility
patterns of these bacterial isolates
towards some traditional antibiotics
were examined due to diameters of
inhibition zones(Tables 2 and 3).
Jar allah et al
Hinton agar plates were prepared and
poured in Petri dishes after sterilization
by autoclaving at 121C for 15 min
.Then it used for testing the isolated
microorganism
for
antibiotic
susceptibility . The diameter of
inhibition zones (mm) were recorded
for all of the plates and then compared
with
the
standard(National
Committee;199).
Results:The results of the present study
showed that thestudied foods of
restaurants were contaminated with
three strains ofpathogenic bacteria,
while thepostive samples of meat
Table 1:Detection of bacterial isolates from meat sample and kebabsample.
Bacterial isolate
Staphylococcus aureus
E.coli
Klebsiellaspp
Gram stain
+
-
Meat samples
40%
29%
-
Kebab samples
28%
19%
9%
30
25
20
15
10
5
0
S. aureus
E. Coli
Klebsiella Spp.
Figure 1: The percent of pathogenic bacteria isolates fromKebab samplesfrom
fast food restaurant.
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Euphrates Journal of Agriculture Science-6 (4): 30-37 , (2014)
Jar allah et al
45
40
35
30
25
20
15
10
5
0
E. Coli
S. aureus
Figure 2:The percent of pathogenic bacteria isolates from meat samples from
butcher shops.
Table 2:Mean of inhibition zone diameters (mm) of bacterial isolates of Kebab
samples.
Antibiotic
Lincomycin
Erythromycin
Tetracyclin
Gentamycin
Amoxicillin
Cefixime
S.aureus
3±0.02
5±0.02
13±0.01
5±0.02
E.coli
Klebsiella sp.
6±0.01
5±0.02
12±0.03
-
12±0.1
7±0.02
8±0.04
12±0.03
5±0.01
6±0.02
Table 3: Means of inhibition zone diameters (mm) of bacterial isolates of meat
samples.
Antibiotics
Lincomycin
Erythromycin
Tetracyclin
Gentamycin
Amoxicillin
Cefixime
(-)mean resistance
S.aureus
E.coli
6±0.01
12±0.02
4±0.02
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10±0.03
6±0.02
7±0.02
13±0.02
6±0.04
5±0.03
Euphrates Journal of Agriculture Science-6 (4): 30-37 , (2014)
isolates may have been the result of
human contamination, a uniform
pattern of human-associated strains was
not observed. This result is agreement
with other reports that S.aureus
frequently are present in low numbers
on raw meat surfaceoccurs infrequently
(Saadiaand Hassanein ,2010).
Biofilm forming bacteria is
usually resistant to a wide range of
antibiotics to find the prevalence of
drug resistance bacteria; assays for
susceptibility profiles and biofilm
formation were performed. Resistance
of bacterial isolates to available
antibiotics and the biofilm formation
ability of these isolates was commonly
observed. The problem may be
attributed to a number of possible
sources,
including
the
natural
resistance of species to certain
antibiotics
possible
transfer
of
antibiotic resistance among species.
The present study demonstrated
that Fast foods samples from
restaurants in Kut City, were heavily
contaminated with S.aureus ( 28% ).
This represent a high level of
contamination indicates a potential
breakdown of hygiene at various stages
of the food processing,S.aureus is most
likely transmitted by hand of food
workers.. Others also reported the
Staphylococcus aureus as a source of
food
contamination
(Kalantariet
al.,2012).
E.coli was the second most
frequent food borne pathogen which
was isolated from Fast foods samples (
19%) ,Food or water or with the
individuals handling the infant's child
and
unwashed
vegetables
or
undercooked meat, by infected food
workers via fecal-oral route. This result
is agreement with other reports, It was
suggested that it could increase the
Discussion:
Meat and meat products are
considered as an excellent source of
high quality animal protein, vitamins
especially B complex, and certain
minerals, especially iron .The source of
infection is not determined in the
majority of food born disease
outbreaks.
Currently
the
most
important pathogens associated with
meat
products
are
Escherichia
coli(Borch et al.,1996).
The present study demonstrated
that meat samples from butchers shops
in Kut City, were heavily contaminated
with E.coli ( 40% ), High
contamination level of Coliforms in
examined meat products may indicates
unsanitary conditions of raw meat
production from which produced. They
are indicators of fecal pollution at
slaughterhouse which begin from
skinning and direct contact with knives
and workers hands. Also, during
evisceration
and
washing,
contamination may come from
intestinal contents as well as from
water during rinsing and washing of
carcasses. Undercooked meat products
have caused many food poisoning
incidents associated with E. coli which
is present in the faeces, intestines and
hide of healthy cattle from where it can
potentially contaminate meat during the
slaughtering process .This result was
remarkably different from those
previously
reported
by(
ElGohary,1993)(78%) and (Vazgeceret
al.,2004) (31%).
In the current study, we
evaluated the prevalence of S. aureus
in meat; the distinct S. aureus
populations on each product type
suggest that meat animals are the
predominant source of contamination.
While a portion of the S. aureus
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Jar allah et al
Euphrates Journal of Agriculture Science-6 (4): 30-37 , (2014)
incidence
of
enteric
pathogens(Angelilloet al.,2000). The
bacterial isolateKlebsiella spp. was the
third most frequent food borne
pathogen which was isolated from Fast
foods samples ( 9%), This is due to the
rapid proliferation of such pathogen in
meat product regardless their count is
high or low.Klebsiellaspp was isolated
in different rates of incidence. As well
as have shown higher incidence
ofKlebsiellasppin
meat
product
(Ammar,2005).
The microbial resistance of
S.aureus isolated from fast food
samples showed a high resistance rate
to commonly used [Lincomycin ,
Erythromycin and amoxicillin ] which
is clinically important and mayindicate
inappropriate use of antimicrobials in
Iraq,while , E. coli isolates showed low
resistance to amoxicillin which is in
agreement with the results of other
investigators in other parts of the world
(Cook et al.,2009).
Klebsiellaspp was relatively
resistant
to
commonly
used
antimicrobials .Our data are in
agreement with several studies in other
parts of the world showing an
increasing portion of resistant isolates
of Klebsiella spp.The prevalence of
antimicrobial resistance among foodborne pathogens has increased during
recent decades.This increase is
attributed to the selection pressure
created by using antimicrobials in
food-producing animals, inaddition to
the unregulated use of antibiotics by
humans
in
developing
countries(Allisonand Gilbert,1995).
Recommendations:- 1- The use of
modern techniques for the diagnosis
isolated bacteria such as ELISA and
PCR. 2-Study of pathological and
histological effects of isolated bacteria
in laboratory animals.
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Jar allah et al
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